Targeted multiplex imaging mass spectrometry with single chain fragment variable (scfv) recombinant antibodies.

Thiery G, Mernaugh RL, Yan H, Spraggins JM, Yang J, Parl FF, Caprioli RM
J Am Soc Mass Spectrom. 2012 23 (10): 1689-96

PMID: 22869296 · PMCID: PMC3525520 · DOI:10.1007/s13361-012-0423-x

Recombinant scfv antibodies specific for CYP1A1 and CYP1B1 P450 enzymes were combined with targeted imaging mass spectrometry to simultaneously detect the P450 enzymes present in archived, paraffin-embedded, human breast cancer tissue sections. By using CYP1A1 and CYP1B1 specific scfv, each coupled to a unique reporter molecule (i.e., a mass tag) it was possible to simultaneously detect multiple antigens within a single tissue sample with high sensitivity and specificity using mass spectrometry. The capability of imaging multiple antigens at the same time is a significant advance that overcomes technical barriers encountered when using present day approaches to develop assays that can simultaneously detect more than a single antigen in the same tissue sample.

MeSH Terms (20)

Animals Antibodies, Monoclonal Aryl Hydrocarbon Hydroxylases Avidin Biotin Breast Breast Neoplasms Cytochrome P-450 CYP1A1 Cytochrome P-450 CYP1B1 Enzyme-Linked Immunosorbent Assay Female Histocytochemistry Humans Mass Spectrometry Mice Molecular Imaging Rats Recombinant Proteins Sensitivity and Specificity Single-Chain Antibodies

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