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The modified SERS active strips were used to capture antigens of interest, incubate Raman probes, and generate SERS maps based on specific Raman peaks of the Raman probes. Simultaneous detection and quantification of multiple biomarkers, including HER2, CA15\u20103, CA27\u201029, PSA, and MUC4, were demonstrated by the immobilization of specific antibodies and the use of uniquely labeled Raman probes. Spatially resolved and co\u2010immobilized multiplexing strategies were developed and successfully validated, showing high specificity and sensitivity with negligible background interference from serum components. 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