<?xml version="1.0" encoding="UTF-8"?>

<!--XBRL Document Created with Wdesk from Workiva-->
<!--Copyright 2023 Workiva-->
<!--r:8ad5ff9b-d21b-4701-8856-2e3503e3299a,g:d6bca12e-7605-4abf-83e7-1fbdbe767d72-->
<link:linkbase xmlns:link="http://www.xbrl.org/2003/linkbase" xmlns:xlink="http://www.w3.org/1999/xlink" xmlns:xsi="http://www.w3.org/2001/XMLSchema-instance" xsi:schemaLocation="http://www.xbrl.org/2003/linkbase http://www.xbrl.org/2003/xbrl-linkbase-2003-12-31.xsd">
  <link:roleRef roleURI="http://www.nautilus.bio/role/CondensedConsolidatedBalanceSheets" xlink:type="simple" xlink:href="naut-20230331.xsd#CondensedConsolidatedBalanceSheets"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CondensedConsolidatedBalanceSheets" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_Liabilities_3d3f4f42-e584-4077-9822-cb087752e98f" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_Liabilities"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilitiesCurrent_bb62ff7c-761c-4181-944c-d8786237ee37" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LiabilitiesCurrent"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_Liabilities_3d3f4f42-e584-4077-9822-cb087752e98f" xlink:to="loc_us-gaap_LiabilitiesCurrent_bb62ff7c-761c-4181-944c-d8786237ee37" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeaseLiabilityNoncurrent_c911eb5b-80d9-49a6-9ce6-be35e61f92eb" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingLeaseLiabilityNoncurrent"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_Liabilities_3d3f4f42-e584-4077-9822-cb087752e98f" xlink:to="loc_us-gaap_OperatingLeaseLiabilityNoncurrent_c911eb5b-80d9-49a6-9ce6-be35e61f92eb" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AssetsCurrent_1db2ccae-3b9b-404f-81ef-db09f9619f0c" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AssetsCurrent"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashAndCashEquivalentsAtCarryingValue_afe64fd4-770c-4d3b-afdc-c9da27187437" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_CashAndCashEquivalentsAtCarryingValue"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_AssetsCurrent_1db2ccae-3b9b-404f-81ef-db09f9619f0c" xlink:to="loc_us-gaap_CashAndCashEquivalentsAtCarryingValue_afe64fd4-770c-4d3b-afdc-c9da27187437" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleSecuritiesDebtSecuritiesCurrent_954897f3-6748-4c17-b553-5c287479ed97" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AvailableForSaleSecuritiesDebtSecuritiesCurrent"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_AssetsCurrent_1db2ccae-3b9b-404f-81ef-db09f9619f0c" xlink:to="loc_us-gaap_AvailableForSaleSecuritiesDebtSecuritiesCurrent_954897f3-6748-4c17-b553-5c287479ed97" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_PrepaidExpenseAndOtherAssetsCurrent_113fc6ad-e0de-43f3-b340-f1d2704b9ff9" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_PrepaidExpenseAndOtherAssetsCurrent"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_AssetsCurrent_1db2ccae-3b9b-404f-81ef-db09f9619f0c" xlink:to="loc_us-gaap_PrepaidExpenseAndOtherAssetsCurrent_113fc6ad-e0de-43f3-b340-f1d2704b9ff9" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_Assets_7eb2ec27-c7c1-43e7-9108-8406ee3d4f57" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_Assets"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AssetsCurrent_88b9024a-29da-48ce-a19b-ea06ac24b554" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AssetsCurrent"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_Assets_7eb2ec27-c7c1-43e7-9108-8406ee3d4f57" xlink:to="loc_us-gaap_AssetsCurrent_88b9024a-29da-48ce-a19b-ea06ac24b554" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentNet_6ac5732f-0238-4d84-a2ed-2baba44044c4" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_PropertyPlantAndEquipmentNet"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_Assets_7eb2ec27-c7c1-43e7-9108-8406ee3d4f57" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentNet_6ac5732f-0238-4d84-a2ed-2baba44044c4" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeaseRightOfUseAsset_adfc5d04-f653-46ac-b35a-fab70519e17c" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingLeaseRightOfUseAsset"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_Assets_7eb2ec27-c7c1-43e7-9108-8406ee3d4f57" xlink:to="loc_us-gaap_OperatingLeaseRightOfUseAsset_adfc5d04-f653-46ac-b35a-fab70519e17c" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherAssetsNoncurrent_ede88739-99c9-4af4-9894-416edcf015f5" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OtherAssetsNoncurrent"/>
    <link:calculationArc order="4" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_Assets_7eb2ec27-c7c1-43e7-9108-8406ee3d4f57" xlink:to="loc_us-gaap_OtherAssetsNoncurrent_ede88739-99c9-4af4-9894-416edcf015f5" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LongTermInvestments_33795b87-e72b-42cc-a46d-cb859a59115f" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LongTermInvestments"/>
    <link:calculationArc order="5" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_Assets_7eb2ec27-c7c1-43e7-9108-8406ee3d4f57" xlink:to="loc_us-gaap_LongTermInvestments_33795b87-e72b-42cc-a46d-cb859a59115f" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockholdersEquity_2ea4c141-a083-4fa8-87a6-9e9dad9308c6" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_StockholdersEquity"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_PreferredStockValue_f0d286bf-fec3-40b9-a7c8-0162cf7e6434" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_PreferredStockValue"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_StockholdersEquity_2ea4c141-a083-4fa8-87a6-9e9dad9308c6" xlink:to="loc_us-gaap_PreferredStockValue_f0d286bf-fec3-40b9-a7c8-0162cf7e6434" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockValue_ca61eb18-1373-4b5f-9cf6-b41310d93a0c" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_CommonStockValue"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_StockholdersEquity_2ea4c141-a083-4fa8-87a6-9e9dad9308c6" xlink:to="loc_us-gaap_CommonStockValue_ca61eb18-1373-4b5f-9cf6-b41310d93a0c" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AdditionalPaidInCapital_5ca2dca0-68c0-4f3a-a461-ee167925e9d4" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AdditionalPaidInCapital"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_StockholdersEquity_2ea4c141-a083-4fa8-87a6-9e9dad9308c6" xlink:to="loc_us-gaap_AdditionalPaidInCapital_5ca2dca0-68c0-4f3a-a461-ee167925e9d4" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccumulatedOtherComprehensiveIncomeLossNetOfTax_3bdbd118-d453-4235-9e69-17a1b1940ffc" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AccumulatedOtherComprehensiveIncomeLossNetOfTax"/>
    <link:calculationArc order="4" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_StockholdersEquity_2ea4c141-a083-4fa8-87a6-9e9dad9308c6" xlink:to="loc_us-gaap_AccumulatedOtherComprehensiveIncomeLossNetOfTax_3bdbd118-d453-4235-9e69-17a1b1940ffc" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_RetainedEarningsAccumulatedDeficit_7cd0dfca-7a86-47ef-9fb2-2b0c3711c189" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_RetainedEarningsAccumulatedDeficit"/>
    <link:calculationArc order="5" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_StockholdersEquity_2ea4c141-a083-4fa8-87a6-9e9dad9308c6" xlink:to="loc_us-gaap_RetainedEarningsAccumulatedDeficit_7cd0dfca-7a86-47ef-9fb2-2b0c3711c189" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilitiesAndStockholdersEquity_9eae7ad2-f0b2-49e0-a7f6-e48a69a40c9d" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LiabilitiesAndStockholdersEquity"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_StockholdersEquity_2b8e7a5c-867e-4f6f-830a-74363968d03a" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_StockholdersEquity"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LiabilitiesAndStockholdersEquity_9eae7ad2-f0b2-49e0-a7f6-e48a69a40c9d" xlink:to="loc_us-gaap_StockholdersEquity_2b8e7a5c-867e-4f6f-830a-74363968d03a" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_Liabilities_8ec68413-2f37-413f-b59e-10862e5060ea" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_Liabilities"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LiabilitiesAndStockholdersEquity_9eae7ad2-f0b2-49e0-a7f6-e48a69a40c9d" xlink:to="loc_us-gaap_Liabilities_8ec68413-2f37-413f-b59e-10862e5060ea" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommitmentsAndContingencies_ab02921f-5e4b-4787-a7d2-a2d2078037d1" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_CommitmentsAndContingencies"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LiabilitiesAndStockholdersEquity_9eae7ad2-f0b2-49e0-a7f6-e48a69a40c9d" xlink:to="loc_us-gaap_CommitmentsAndContingencies_ab02921f-5e4b-4787-a7d2-a2d2078037d1" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LiabilitiesCurrent_368e8461-d6fe-44aa-bf9b-6f3f711953a0" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LiabilitiesCurrent"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeaseLiabilityCurrent_3964f329-1c10-4be8-a128-891c0f39646f" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingLeaseLiabilityCurrent"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LiabilitiesCurrent_368e8461-d6fe-44aa-bf9b-6f3f711953a0" xlink:to="loc_us-gaap_OperatingLeaseLiabilityCurrent_3964f329-1c10-4be8-a128-891c0f39646f" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent_43b1defe-2a0e-4fc8-af08-3deb17ab8862" xlink:href="naut-20230331.xsd#naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LiabilitiesCurrent_368e8461-d6fe-44aa-bf9b-6f3f711953a0" xlink:to="loc_naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent_43b1defe-2a0e-4fc8-af08-3deb17ab8862" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccountsPayableCurrent_fd4817f3-cf26-4dbd-b810-cae069cacd64" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AccountsPayableCurrent"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LiabilitiesCurrent_368e8461-d6fe-44aa-bf9b-6f3f711953a0" xlink:to="loc_us-gaap_AccountsPayableCurrent_fd4817f3-cf26-4dbd-b810-cae069cacd64" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CondensedConsolidatedStatementsofOperations" xlink:type="simple" xlink:href="naut-20230331.xsd#CondensedConsolidatedStatementsofOperations"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CondensedConsolidatedStatementsofOperations" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NonoperatingIncomeExpense_670f748d-95d0-49db-81e1-48332f472be2" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NonoperatingIncomeExpense"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherNonoperatingExpense_df18e0ce-e86b-47e9-8755-02d2ced3b6fc" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OtherNonoperatingExpense"/>
    <link:calculationArc order="1" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NonoperatingIncomeExpense_670f748d-95d0-49db-81e1-48332f472be2" xlink:to="loc_us-gaap_OtherNonoperatingExpense_df18e0ce-e86b-47e9-8755-02d2ced3b6fc" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_InterestIncomeExpenseNonoperatingNet_ba824912-bc16-41f5-91f2-db110b20c489" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_InterestIncomeExpenseNonoperatingNet"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NonoperatingIncomeExpense_670f748d-95d0-49db-81e1-48332f472be2" xlink:to="loc_us-gaap_InterestIncomeExpenseNonoperatingNet_ba824912-bc16-41f5-91f2-db110b20c489" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingExpenses_569c7314-93c0-413c-8a2d-1a02b1ff99ca" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingExpenses"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_GeneralAndAdministrativeExpense_deb5c59a-6fdc-44bb-803d-c6a7a857c934" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_GeneralAndAdministrativeExpense"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_OperatingExpenses_569c7314-93c0-413c-8a2d-1a02b1ff99ca" xlink:to="loc_us-gaap_GeneralAndAdministrativeExpense_deb5c59a-6fdc-44bb-803d-c6a7a857c934" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ResearchAndDevelopmentExpense_f51c9117-5d57-40a4-bec3-e51e06828735" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ResearchAndDevelopmentExpense"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_OperatingExpenses_569c7314-93c0-413c-8a2d-1a02b1ff99ca" xlink:to="loc_us-gaap_ResearchAndDevelopmentExpense_f51c9117-5d57-40a4-bec3-e51e06828735" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetIncomeLoss_25631710-4608-44e6-935a-1fd9e5a1e8da" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetIncomeLoss"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingExpenses_bef01372-0e6f-4cca-a12c-3015f59ab099" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingExpenses"/>
    <link:calculationArc order="1" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetIncomeLoss_25631710-4608-44e6-935a-1fd9e5a1e8da" xlink:to="loc_us-gaap_OperatingExpenses_bef01372-0e6f-4cca-a12c-3015f59ab099" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NonoperatingIncomeExpense_9441ffd9-840f-4dde-81af-272a33d318d1" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NonoperatingIncomeExpense"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetIncomeLoss_25631710-4608-44e6-935a-1fd9e5a1e8da" xlink:to="loc_us-gaap_NonoperatingIncomeExpense_9441ffd9-840f-4dde-81af-272a33d318d1" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CondensedConsolidatedStatementsofComprehensiveLoss" xlink:type="simple" xlink:href="naut-20230331.xsd#CondensedConsolidatedStatementsofComprehensiveLoss"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CondensedConsolidatedStatementsofComprehensiveLoss" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ComprehensiveIncomeNetOfTax_3195bff8-f114-421d-bd1c-b240a238c87e" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ComprehensiveIncomeNetOfTax"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_231ee23e-1d46-4c2d-b5b6-2db4191ab54e" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_ComprehensiveIncomeNetOfTax_3195bff8-f114-421d-bd1c-b240a238c87e" xlink:to="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_231ee23e-1d46-4c2d-b5b6-2db4191ab54e" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetIncomeLoss_707e2513-da3d-44ef-97e6-a8776b228670" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetIncomeLoss"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_ComprehensiveIncomeNetOfTax_3195bff8-f114-421d-bd1c-b240a238c87e" xlink:to="loc_us-gaap_NetIncomeLoss_707e2513-da3d-44ef-97e6-a8776b228670" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_ce1bf1cb-05a2-4377-a6bc-484ec17db4cd" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherComprehensiveIncomeAvailableforsaleSecuritiesAdjustmentNetOfTaxPortionAttributableToParent_d106ff81-e594-4440-9c6d-254283bbb07b" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OtherComprehensiveIncomeAvailableforsaleSecuritiesAdjustmentNetOfTaxPortionAttributableToParent"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_OtherComprehensiveIncomeLossNetOfTaxPortionAttributableToParent_ce1bf1cb-05a2-4377-a6bc-484ec17db4cd" xlink:to="loc_us-gaap_OtherComprehensiveIncomeAvailableforsaleSecuritiesAdjustmentNetOfTaxPortionAttributableToParent_d106ff81-e594-4440-9c6d-254283bbb07b" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CondensedConsolidatedStatementsofCashFlows" xlink:type="simple" xlink:href="naut-20230331.xsd#CondensedConsolidatedStatementsofCashFlows"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CondensedConsolidatedStatementsofCashFlows" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalentsPeriodIncreaseDecreaseIncludingExchangeRateEffect_513fd6c0-8cdd-493b-abce-089d127b880e" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalentsPeriodIncreaseDecreaseIncludingExchangeRateEffect"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_946827a1-b4fe-4e70-ad7a-77b9f4b1bc6c" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetCashProvidedByUsedInInvestingActivities"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalentsPeriodIncreaseDecreaseIncludingExchangeRateEffect_513fd6c0-8cdd-493b-abce-089d127b880e" xlink:to="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_946827a1-b4fe-4e70-ad7a-77b9f4b1bc6c" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_71655302-ac8e-4fb7-81df-d4c435d84f5b" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetCashProvidedByUsedInOperatingActivities"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalentsPeriodIncreaseDecreaseIncludingExchangeRateEffect_513fd6c0-8cdd-493b-abce-089d127b880e" xlink:to="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_71655302-ac8e-4fb7-81df-d4c435d84f5b" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInFinancingActivities_7f4d57bd-1424-4ed2-830e-9a253c9ee092" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetCashProvidedByUsedInFinancingActivities"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalentsPeriodIncreaseDecreaseIncludingExchangeRateEffect_513fd6c0-8cdd-493b-abce-089d127b880e" xlink:to="loc_us-gaap_NetCashProvidedByUsedInFinancingActivities_7f4d57bd-1424-4ed2-830e-9a253c9ee092" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetCashProvidedByUsedInOperatingActivities"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccretionAmortizationOfDiscountsAndPremiumsInvestments_93d13426-1f5e-4fe9-9a66-3778dbaf5f2b" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AccretionAmortizationOfDiscountsAndPremiumsInvestments"/>
    <link:calculationArc order="1" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_AccretionAmortizationOfDiscountsAndPremiumsInvestments_93d13426-1f5e-4fe9-9a66-3778dbaf5f2b" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetIncomeLoss_2b8efbef-40f0-4c9d-ae96-03af04a295c8" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetIncomeLoss"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_NetIncomeLoss_2b8efbef-40f0-4c9d-ae96-03af04a295c8" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncreaseDecreaseInOperatingLeaseLiability_9b813e87-3c96-430c-8410-a365c99af87f" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_IncreaseDecreaseInOperatingLeaseLiability"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_IncreaseDecreaseInOperatingLeaseLiability_9b813e87-3c96-430c-8410-a365c99af87f" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeaseRightOfUseAssetAmortizationExpense_efadf0a3-a06d-490b-9959-ec2180d541d6" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingLeaseRightOfUseAssetAmortizationExpense"/>
    <link:calculationArc order="4" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_OperatingLeaseRightOfUseAssetAmortizationExpense_efadf0a3-a06d-490b-9959-ec2180d541d6" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensation_15fd5b56-5336-4186-ba76-ed5960bbbb89" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ShareBasedCompensation"/>
    <link:calculationArc order="5" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_ShareBasedCompensation_15fd5b56-5336-4186-ba76-ed5960bbbb89" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncreaseDecreaseInAccountsPayable_c321bc9b-689d-412d-a5ad-338d49d53735" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_IncreaseDecreaseInAccountsPayable"/>
    <link:calculationArc order="6" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_IncreaseDecreaseInAccountsPayable_c321bc9b-689d-412d-a5ad-338d49d53735" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_Depreciation_4077a2bf-43f4-4878-928c-2266c48d8fb9" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_Depreciation"/>
    <link:calculationArc order="7" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_Depreciation_4077a2bf-43f4-4878-928c-2266c48d8fb9" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncreaseDecreaseInPrepaidDeferredExpenseAndOtherAssets_cc801990-ba86-43ce-841f-2a69c3b679c0" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_IncreaseDecreaseInPrepaidDeferredExpenseAndOtherAssets"/>
    <link:calculationArc order="8" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_IncreaseDecreaseInPrepaidDeferredExpenseAndOtherAssets_cc801990-ba86-43ce-841f-2a69c3b679c0" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_IncreaseDecreaseInAccruedLiabilitiesAndOtherOperatingLiabilities_6208be91-f2ab-46b1-b53e-7d8b370e5f22" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_IncreaseDecreaseInAccruedLiabilitiesAndOtherOperatingLiabilities"/>
    <link:calculationArc order="9" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInOperatingActivities_0e5f21ea-6b07-4716-b5ac-f0e7b9737d94" xlink:to="loc_us-gaap_IncreaseDecreaseInAccruedLiabilitiesAndOtherOperatingLiabilities_6208be91-f2ab-46b1-b53e-7d8b370e5f22" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInFinancingActivities_4d93fe21-de13-4765-9c59-1b3ac60c563e" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetCashProvidedByUsedInFinancingActivities"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromStockOptionsExercised_2e173666-a85b-402e-9895-e3737161f130" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ProceedsFromStockOptionsExercised"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInFinancingActivities_4d93fe21-de13-4765-9c59-1b3ac60c563e" xlink:to="loc_us-gaap_ProceedsFromStockOptionsExercised_2e173666-a85b-402e-9895-e3737161f130" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_c329bf24-ae77-4222-9684-01426870e090" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_NetCashProvidedByUsedInInvestingActivities"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ProceedsFromSaleAndMaturityOfAvailableForSaleSecurities_8808de2f-77df-49e5-ac94-d2f9649895e8" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ProceedsFromSaleAndMaturityOfAvailableForSaleSecurities"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_c329bf24-ae77-4222-9684-01426870e090" xlink:to="loc_us-gaap_ProceedsFromSaleAndMaturityOfAvailableForSaleSecurities_8808de2f-77df-49e5-ac94-d2f9649895e8" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsToAcquirePropertyPlantAndEquipment_a576bf15-fe86-4694-a3a4-74a8fc2a4989" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_PaymentsToAcquirePropertyPlantAndEquipment"/>
    <link:calculationArc order="2" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_c329bf24-ae77-4222-9684-01426870e090" xlink:to="loc_us-gaap_PaymentsToAcquirePropertyPlantAndEquipment_a576bf15-fe86-4694-a3a4-74a8fc2a4989" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_PaymentsToAcquireAvailableForSaleSecuritiesDebt_690dec39-3ee5-4692-8ccb-4fca38bf72ac" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_PaymentsToAcquireAvailableForSaleSecuritiesDebt"/>
    <link:calculationArc order="3" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_NetCashProvidedByUsedInInvestingActivities_c329bf24-ae77-4222-9684-01426870e090" xlink:to="loc_us-gaap_PaymentsToAcquireAvailableForSaleSecuritiesDebt_690dec39-3ee5-4692-8ccb-4fca38bf72ac" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/FairValueMeasurementsAssetsCarriedatFairValueandMeasuredonaRecurringBasisDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#FairValueMeasurementsAssetsCarriedatFairValueandMeasuredonaRecurringBasisDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/FairValueMeasurementsAssetsCarriedatFairValueandMeasuredonaRecurringBasisDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleDebtSecuritiesAmortizedCostBasis_c7974928-d352-4e73-8d06-fbd66e3c8a51" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AvailableForSaleDebtSecuritiesAmortizedCostBasis"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleDebtSecuritiesAccumulatedGrossUnrealizedGainBeforeTax_7a4873e9-58ac-454f-9ba3-5f1b16a6241d" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AvailableForSaleDebtSecuritiesAccumulatedGrossUnrealizedGainBeforeTax"/>
    <link:calculationArc order="1" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_AvailableForSaleDebtSecuritiesAmortizedCostBasis_c7974928-d352-4e73-8d06-fbd66e3c8a51" xlink:to="loc_us-gaap_AvailableForSaleDebtSecuritiesAccumulatedGrossUnrealizedGainBeforeTax_7a4873e9-58ac-454f-9ba3-5f1b16a6241d" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleSecuritiesDebtSecurities_fbbac39c-5fa2-4b4b-b8dd-52ad4241442d" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AvailableForSaleSecuritiesDebtSecurities"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_AvailableForSaleDebtSecuritiesAmortizedCostBasis_c7974928-d352-4e73-8d06-fbd66e3c8a51" xlink:to="loc_us-gaap_AvailableForSaleSecuritiesDebtSecurities_fbbac39c-5fa2-4b4b-b8dd-52ad4241442d" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AvailableForSaleDebtSecuritiesAccumulatedGrossUnrealizedLossBeforeTax_74f0cc9e-359a-4808-a1fb-e98f63d964f7" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AvailableForSaleDebtSecuritiesAccumulatedGrossUnrealizedLossBeforeTax"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_AvailableForSaleDebtSecuritiesAmortizedCostBasis_c7974928-d352-4e73-8d06-fbd66e3c8a51" xlink:to="loc_us-gaap_AvailableForSaleDebtSecuritiesAccumulatedGrossUnrealizedLossBeforeTax_74f0cc9e-359a-4808-a1fb-e98f63d964f7" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/FairValueMeasurementsUnrealizedLossesandFairValuesofAvailableforSaleSecuritiesinanUnrealizedLossPositionDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#FairValueMeasurementsUnrealizedLossesandFairValuesofAvailableforSaleSecuritiesinanUnrealizedLossPositionDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/FairValueMeasurementsUnrealizedLossesandFairValuesofAvailableforSaleSecuritiesinanUnrealizedLossPositionDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPositionAccumulatedLoss_cd330986-c13d-4c2f-80e0-89029ad3752f" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPositionAccumulatedLoss"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPositionLessThan12MonthsAccumulatedLoss_568d13cc-667a-4cef-b8d0-3d8d2bf118d6" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPositionLessThan12MonthsAccumulatedLoss"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPositionAccumulatedLoss_cd330986-c13d-4c2f-80e0-89029ad3752f" xlink:to="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPositionLessThan12MonthsAccumulatedLoss_568d13cc-667a-4cef-b8d0-3d8d2bf118d6" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPosition12MonthsOrLongerAccumulatedLoss_73fd3f03-e810-4c35-8a97-e92aeb1c8092" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPosition12MonthsOrLongerAccumulatedLoss"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPositionAccumulatedLoss_cd330986-c13d-4c2f-80e0-89029ad3752f" xlink:to="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPosition12MonthsOrLongerAccumulatedLoss_73fd3f03-e810-4c35-8a97-e92aeb1c8092" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPosition_0bac9e33-6376-41bc-913b-398d100ff7a3" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPosition"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPosition12MonthsOrLonger_f59fdf75-8d8f-4cae-8bf7-6477f68d617c" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPosition12MonthsOrLonger"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPosition_0bac9e33-6376-41bc-913b-398d100ff7a3" xlink:to="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPosition12MonthsOrLonger_f59fdf75-8d8f-4cae-8bf7-6477f68d617c" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPositionLessThan12Months_30aff46e-712f-4aed-ac2c-84464d91cdab" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPositionLessThan12Months"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_DebtSecuritiesAvailableForSaleUnrealizedLossPosition_0bac9e33-6376-41bc-913b-398d100ff7a3" xlink:to="loc_us-gaap_DebtSecuritiesAvailableForSaleContinuousUnrealizedLossPositionLessThan12Months_30aff46e-712f-4aed-ac2c-84464d91cdab" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsPropertyandEquipmentNetDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsPropertyandEquipmentNetDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsPropertyandEquipmentNetDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentNet_437f12ee-1f50-43d2-884b-2fefe577c7a4" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_PropertyPlantAndEquipmentNet"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment_f0482ad3-ccbb-42df-ae02-037d25f1b26e" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment"/>
    <link:calculationArc order="1" weight="-1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentNet_437f12ee-1f50-43d2-884b-2fefe577c7a4" xlink:to="loc_us-gaap_AccumulatedDepreciationDepletionAndAmortizationPropertyPlantAndEquipment_f0482ad3-ccbb-42df-ae02-037d25f1b26e" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_PropertyPlantAndEquipmentGross_a0945700-2d8c-441d-b010-90bd6f8d6304" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_PropertyPlantAndEquipmentGross"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_PropertyPlantAndEquipmentNet_437f12ee-1f50-43d2-884b-2fefe577c7a4" xlink:to="loc_us-gaap_PropertyPlantAndEquipmentGross_a0945700-2d8c-441d-b010-90bd6f8d6304" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsOtherLongtermAssetsDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsOtherLongtermAssetsDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsOtherLongtermAssetsDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherAssetsNoncurrent_25b7f347-7f79-4290-ba8c-cb21312219a3" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OtherAssetsNoncurrent"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_DepositsAssetsNoncurrent_52694eb2-dea6-48e1-8ba7-6dfd884c6706" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_DepositsAssetsNoncurrent"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_OtherAssetsNoncurrent_25b7f347-7f79-4290-ba8c-cb21312219a3" xlink:to="loc_us-gaap_DepositsAssetsNoncurrent_52694eb2-dea6-48e1-8ba7-6dfd884c6706" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedCashNoncurrent_7a6b2ee4-fec0-4cc9-8597-00da53ea3c68" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_RestrictedCashNoncurrent"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_OtherAssetsNoncurrent_25b7f347-7f79-4290-ba8c-cb21312219a3" xlink:to="loc_us-gaap_RestrictedCashNoncurrent_7a6b2ee4-fec0-4cc9-8597-00da53ea3c68" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsAccruedExpensesandOtherLiabilitiesDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsAccruedExpensesandOtherLiabilitiesDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsAccruedExpensesandOtherLiabilitiesDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent_ec180815-4b89-4484-baf1-76fae7c1b620" xlink:href="naut-20230331.xsd#naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent"/>
    <link:loc xlink:type="locator" xlink:label="loc_naut_AccruedResearchAndDevelopment_f143b120-7fbe-4752-81b4-9998dfc57276" xlink:href="naut-20230331.xsd#naut_AccruedResearchAndDevelopment"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent_ec180815-4b89-4484-baf1-76fae7c1b620" xlink:to="loc_naut_AccruedResearchAndDevelopment_f143b120-7fbe-4752-81b4-9998dfc57276" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_EmployeeRelatedLiabilitiesCurrent_0e2de427-c9b5-4c98-a9b5-e28cdbe50a9e" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_EmployeeRelatedLiabilitiesCurrent"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent_ec180815-4b89-4484-baf1-76fae7c1b620" xlink:to="loc_us-gaap_EmployeeRelatedLiabilitiesCurrent_0e2de427-c9b5-4c98-a9b5-e28cdbe50a9e" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OtherSundryLiabilitiesCurrent_d37f5eb6-87b3-4f87-95d5-3eedae45cbe0" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OtherSundryLiabilitiesCurrent"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent_ec180815-4b89-4484-baf1-76fae7c1b620" xlink:to="loc_us-gaap_OtherSundryLiabilitiesCurrent_d37f5eb6-87b3-4f87-95d5-3eedae45cbe0" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_AccruedProfessionalFeesCurrent_eee12077-9ef3-4f61-9e43-39db18b00b9a" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_AccruedProfessionalFeesCurrent"/>
    <link:calculationArc order="4" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_naut_AccruedLiabilitiesAndOtherLiabilitiesCurrent_ec180815-4b89-4484-baf1-76fae7c1b620" xlink:to="loc_us-gaap_AccruedProfessionalFeesCurrent_eee12077-9ef3-4f61-9e43-39db18b00b9a" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsCashCashEquivalentsandRestrictedCashDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsCashCashEquivalentsandRestrictedCashDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CompositionofCertainCondensedConsolidatedFinancialStatementLineItemsCashCashEquivalentsandRestrictedCashDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalents_5c23d612-7de7-4b94-bbea-7377a12b5f68" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalents"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_RestrictedCashAndCashEquivalents_7a2282bc-a709-45b3-b70e-cefe985f9414" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_RestrictedCashAndCashEquivalents"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalents_5c23d612-7de7-4b94-bbea-7377a12b5f68" xlink:to="loc_us-gaap_RestrictedCashAndCashEquivalents_7a2282bc-a709-45b3-b70e-cefe985f9414" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_CashAndCashEquivalentsAtCarryingValue_db160d00-1e0d-47b1-9c1e-33f40071d029" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_CashAndCashEquivalentsAtCarryingValue"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_CashCashEquivalentsRestrictedCashAndRestrictedCashEquivalents_5c23d612-7de7-4b94-bbea-7377a12b5f68" xlink:to="loc_us-gaap_CashAndCashEquivalentsAtCarryingValue_db160d00-1e0d-47b1-9c1e-33f40071d029" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CommonStockCommonStockReservedforFutureIssuanceDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#CommonStockCommonStockReservedforFutureIssuanceDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CommonStockCommonStockReservedforFutureIssuanceDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_CommonStockCapitalSharesReservedForFutureIssuance_9c5f5cfe-9ecd-47bf-8957-17b1671f2144" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_CommonStockCapitalSharesReservedForFutureIssuance"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNumberOfSharesAvailableForGrant_486ee9a5-dd32-4c06-8f0f-87f7c5c08d12" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNumberOfSharesAvailableForGrant"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_CommonStockCapitalSharesReservedForFutureIssuance_9c5f5cfe-9ecd-47bf-8957-17b1671f2144" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardNumberOfSharesAvailableForGrant_486ee9a5-dd32-4c06-8f0f-87f7c5c08d12" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingNumber_c0f950b0-6207-4dfc-b631-e1086dd964db" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingNumber"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_CommonStockCapitalSharesReservedForFutureIssuance_9c5f5cfe-9ecd-47bf-8957-17b1671f2144" xlink:to="loc_us-gaap_ShareBasedCompensationArrangementByShareBasedPaymentAwardOptionsOutstandingNumber_c0f950b0-6207-4dfc-b631-e1086dd964db" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CommitmentandContingenciesComponentsofLeaseCostDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#CommitmentandContingenciesComponentsofLeaseCostDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CommitmentandContingenciesComponentsofLeaseCostDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LeaseCost_0a952087-2fc2-48ed-9503-6ec722080697" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LeaseCost"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeaseCost_5afd4f42-0b31-45aa-b9b2-c70a191a5ad6" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingLeaseCost"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LeaseCost_0a952087-2fc2-48ed-9503-6ec722080697" xlink:to="loc_us-gaap_OperatingLeaseCost_5afd4f42-0b31-45aa-b9b2-c70a191a5ad6" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_VariableLeaseCost_dc8600bd-70f7-47a1-bf52-95c46d38f0a3" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_VariableLeaseCost"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LeaseCost_0a952087-2fc2-48ed-9503-6ec722080697" xlink:to="loc_us-gaap_VariableLeaseCost_dc8600bd-70f7-47a1-bf52-95c46d38f0a3" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_ShortTermLeaseCost_50f2dfb7-b36f-4c82-b8dd-767d5158ead8" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_ShortTermLeaseCost"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LeaseCost_0a952087-2fc2-48ed-9503-6ec722080697" xlink:to="loc_us-gaap_ShortTermLeaseCost_50f2dfb7-b36f-4c82-b8dd-767d5158ead8" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CommitmentandContingenciesLeaseMaturityDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#CommitmentandContingenciesLeaseMaturityDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CommitmentandContingenciesLeaseMaturityDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_f61ac847-ffb3-467d-95bf-b9a7841b59c1" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearFour_434a84ab-d5f0-4f26-a3cb-0b3b964d26d6" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearFour"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_f61ac847-ffb3-467d-95bf-b9a7841b59c1" xlink:to="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearFour_434a84ab-d5f0-4f26-a3cb-0b3b964d26d6" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearThree_493b44bd-7072-4eb2-bd2d-5d32341c08cb" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearThree"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_f61ac847-ffb3-467d-95bf-b9a7841b59c1" xlink:to="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearThree_493b44bd-7072-4eb2-bd2d-5d32341c08cb" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsRemainderOfFiscalYear_6bfe6f2e-73ff-4a6a-87fb-4a96c34772bb" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityPaymentsRemainderOfFiscalYear"/>
    <link:calculationArc order="3" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_f61ac847-ffb3-467d-95bf-b9a7841b59c1" xlink:to="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsRemainderOfFiscalYear_6bfe6f2e-73ff-4a6a-87fb-4a96c34772bb" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearTwo_2d5cf4d9-1058-411e-b960-08766b5a2557" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearTwo"/>
    <link:calculationArc order="4" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_f61ac847-ffb3-467d-95bf-b9a7841b59c1" xlink:to="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueYearTwo_2d5cf4d9-1058-411e-b960-08766b5a2557" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_naut_LesseeOperatingLeaseLiabilityToBePaidDueAfterYearFour_6aff730f-5743-4db1-adc6-ff00791a15b4" xlink:href="naut-20230331.xsd#naut_LesseeOperatingLeaseLiabilityToBePaidDueAfterYearFour"/>
    <link:calculationArc order="5" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_f61ac847-ffb3-467d-95bf-b9a7841b59c1" xlink:to="loc_naut_LesseeOperatingLeaseLiabilityToBePaidDueAfterYearFour_6aff730f-5743-4db1-adc6-ff00791a15b4" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueNextTwelveMonths_13e7834f-1fab-47cc-8370-3b588e50566f" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueNextTwelveMonths"/>
    <link:calculationArc order="6" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_f61ac847-ffb3-467d-95bf-b9a7841b59c1" xlink:to="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDueNextTwelveMonths_13e7834f-1fab-47cc-8370-3b588e50566f" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/CommitmentandContingenciesLeaseMaturityDetails_1" xlink:type="simple" xlink:href="naut-20230331.xsd#CommitmentandContingenciesLeaseMaturityDetails_1"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/CommitmentandContingenciesLeaseMaturityDetails_1" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_11507bf7-29b4-4b9a-bb0b-2ed001d543c9" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_LesseeOperatingLeaseLiabilityUndiscountedExcessAmount_b27012ee-5203-4752-9d41-12cbf7005527" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_LesseeOperatingLeaseLiabilityUndiscountedExcessAmount"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_11507bf7-29b4-4b9a-bb0b-2ed001d543c9" xlink:to="loc_us-gaap_LesseeOperatingLeaseLiabilityUndiscountedExcessAmount_b27012ee-5203-4752-9d41-12cbf7005527" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_OperatingLeaseLiability_758efe50-1fb3-49f8-8ca2-1f73f9d7bb89" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_OperatingLeaseLiability"/>
    <link:calculationArc order="2" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_LesseeOperatingLeaseLiabilityPaymentsDue_11507bf7-29b4-4b9a-bb0b-2ed001d543c9" xlink:to="loc_us-gaap_OperatingLeaseLiability_758efe50-1fb3-49f8-8ca2-1f73f9d7bb89" xlink:type="arc"/>
  </link:calculationLink>
  <link:roleRef roleURI="http://www.nautilus.bio/role/BasicandDilutedNetLossperShareBasicandDilutedNotLossperShareDetails" xlink:type="simple" xlink:href="naut-20230331.xsd#BasicandDilutedNetLossperShareBasicandDilutedNotLossperShareDetails"/>
  <link:calculationLink xlink:role="http://www.nautilus.bio/role/BasicandDilutedNetLossperShareBasicandDilutedNotLossperShareDetails" xlink:type="extended">
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_WeightedAverageNumberOfSharesOutstandingBasic_8b9116ed-d131-490d-aab0-7283d6e95ff9" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_WeightedAverageNumberOfSharesOutstandingBasic"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_WeightedAverageNumberOfSharesIssuedBasic_5e39e8f5-0f32-482f-997f-6a4368b42b50" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_WeightedAverageNumberOfSharesIssuedBasic"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_WeightedAverageNumberOfSharesOutstandingBasic_8b9116ed-d131-490d-aab0-7283d6e95ff9" xlink:to="loc_us-gaap_WeightedAverageNumberOfSharesIssuedBasic_5e39e8f5-0f32-482f-997f-6a4368b42b50" xlink:type="arc"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_WeightedAverageNumberOfDilutedSharesOutstanding_fefd61f9-bc53-4dbf-b057-fca14f44b67d" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_WeightedAverageNumberOfDilutedSharesOutstanding"/>
    <link:loc xlink:type="locator" xlink:label="loc_us-gaap_WeightedAverageNumberOfSharesIssuedBasic_914b1621-699b-4681-a41e-6cf71787114d" xlink:href="https://xbrl.fasb.org/us-gaap/2022/elts/us-gaap-2022.xsd#us-gaap_WeightedAverageNumberOfSharesIssuedBasic"/>
    <link:calculationArc order="1" weight="1.0" xlink:arcrole="http://www.xbrl.org/2003/arcrole/summation-item" xlink:from="loc_us-gaap_WeightedAverageNumberOfDilutedSharesOutstanding_fefd61f9-bc53-4dbf-b057-fca14f44b67d" xlink:to="loc_us-gaap_WeightedAverageNumberOfSharesIssuedBasic_914b1621-699b-4681-a41e-6cf71787114d" xlink:type="arc"/>
  </link:calculationLink>
</link:linkbase>
