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

    <schemaVersion>X0508</schemaVersion>

    <documentType>4</documentType>

    <periodOfReport>2023-09-27</periodOfReport>

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

    <reportingOwner>
        <reportingOwnerId>
            <rptOwnerCik>0001926254</rptOwnerCik>
            <rptOwnerName>Hackman Jeffrey S.</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>1</isOfficer>
            <isTenPercentOwner>0</isTenPercentOwner>
            <isOther>0</isOther>
            <officerTitle>See Remarks.</officerTitle>
        </reportingOwnerRelationship>
    </reportingOwner>

    <aff10b5One>0</aff10b5One>

    <derivativeTable>
        <derivativeTransaction>
            <securityTitle>
                <value>Stock Option (Right to Purchase)</value>
            </securityTitle>
            <conversionOrExercisePrice>
                <value>0.376</value>
            </conversionOrExercisePrice>
            <transactionDate>
                <value>2023-09-27</value>
            </transactionDate>
            <transactionCoding>
                <transactionFormType>4</transactionFormType>
                <transactionCode>A</transactionCode>
                <equitySwapInvolved>0</equitySwapInvolved>
            </transactionCoding>
            <transactionAmounts>
                <transactionShares>
                    <value>600000</value>
                </transactionShares>
                <transactionPricePerShare>
                    <value>0</value>
                </transactionPricePerShare>
                <transactionAcquiredDisposedCode>
                    <value>A</value>
                </transactionAcquiredDisposedCode>
            </transactionAmounts>
            <exerciseDate>
                <footnoteId id="F1"/>
            </exerciseDate>
            <expirationDate>
                <value>2033-09-27</value>
            </expirationDate>
            <underlyingSecurity>
                <underlyingSecurityTitle>
                    <value>Common Stock</value>
                </underlyingSecurityTitle>
                <underlyingSecurityShares>
                    <value>600000</value>
                </underlyingSecurityShares>
            </underlyingSecurity>
            <postTransactionAmounts>
                <sharesOwnedFollowingTransaction>
                    <value>600000</value>
                </sharesOwnedFollowingTransaction>
            </postTransactionAmounts>
            <ownershipNature>
                <directOrIndirectOwnership>
                    <value>D</value>
                </directOrIndirectOwnership>
            </ownershipNature>
        </derivativeTransaction>
    </derivativeTable>

    <footnotes>
        <footnote id="F1">25% of the shares underlying the stock option vest on September 27, 2024, with the remaining shares vesting on a monthly basis on the 27th day of each month thereafter, in each case, subject to continuous service to the issuer, such that the option shall be fully vested on September 27, 2027.</footnote>
    </footnotes>

    <remarks>Chairman, President, and Chief Executive Officer.</remarks>

    <ownerSignature>
        <signatureName>/s/ Ryan M. Rourke Reed, Attorney-in-Fact</signatureName>
        <signatureDate>2023-09-28</signatureDate>
    </ownerSignature>
</ownershipDocument>
