<?xml version="1.0"?>
<ownershipDocument>

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2023-11-09</periodOfReport>

    <issuer>
        <issuerCik>0001907685</issuerCik>
        <issuerName>Comera Life Sciences Holdings, Inc.</issuerName>
        <issuerTradingSymbol>CMRA</issuerTradingSymbol>
    </issuer>

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001926535</rptOwnerCik>
            <rptOwnerName>Banerjee Sirshendu Roopom</rptOwnerName>
        </reportingOwnerId>
        <reportingOwnerAddress>
            <rptOwnerStreet1>C/O COMERA LIFE SCIENCES HOLDINGS, INC.</rptOwnerStreet1>
            <rptOwnerStreet2>12 GILL STREET, SUITE 4650</rptOwnerStreet2>
            <rptOwnerCity>WOBURN</rptOwnerCity>
            <rptOwnerState>MA</rptOwnerState>
            <rptOwnerZipCode>01801</rptOwnerZipCode>
            <rptOwnerStateDescription></rptOwnerStateDescription>
        </reportingOwnerAddress>
        <reportingOwnerRelationship>
            <isDirector>1</isDirector>
            <isOfficer>0</isOfficer>
            <isTenPercentOwner>0</isTenPercentOwner>
            <isOther>0</isOther>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <derivativeTable>
        <derivativeTransaction>
            <securityTitle>
                <value>Stock Option (Right to Purchase)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>0.3204</value>
            </conversionOrExercisePrice>
            <transactionDate>
                <value>2023-11-09</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>A</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>122487</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>0</value>
                    <footnoteId id="F2"/>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <value>2023-11-09</value>
                <footnoteId id="F1"/>
            </exerciseDate>
            <expirationDate>
                <value>2033-11-09</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>122487</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>122487</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeTransaction>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">Granted pursuant to the Comera Life Sciences Holdings, Inc. 2022 Equity and Incentive Plan. The options were fully vested on the date of grant.</footnote>
        <footnote id="F2">In furtherance of the registrant's cash conservation efforts, the options were granted in lieu of the annual cash retainer and committee fees due to non-employee directors for the quarter ended September 30, 2023, in accordance with the registrant's February 2023 temporary modification to its non-employee director compensation plan. This option was issued to the reporting person in lieu of retainer fees of $16,250. For more information related to the registrant's non-employee director compensation plan and the February 2023 temporary modification, see the registrant's Annual Report on Form 10-K for the year ended December 31, 2022.</footnote>
    </footnotes>

    <ownerSignature>
        <signatureName>/s/ Ryan M. Rourke Reed, Attorney-in-Fact</signatureName>
        <signatureDate>2023-11-14</signatureDate>
    </ownerSignature>
</ownershipDocument>
