Rapid, Sensitive and Quantitative Methods for Tissue and Cell-Based Proteomics Via Consecutive Additional Quantifiable Extenders

Abstract

Methods, systems and kits are provided for detecting and quantifying multiple immunogens in a sample via consecutive addition of quantifiable extenders to immunogen bound complexes of immunogen binding agent attached to a DNA containing an RNA polymerase promoter.

1 . An epitope detector comprising an antibody, a single chain Fv, or a constrained epitope specific complementarity determining region (CDR) for a selected epitope, modified for attachment of an oligonucleotide at a single site. 
     
     
         2 . A method for detecting and quantifying multiple immunogens in a sample comprising:
 (a) forming immunogen binding agent-DNA-oligo complexes via a plurality of epitope detectors of  claim 1  by contacting an immobilized form of immunogens with the plurality of epitope detectors, wherein the DNA of the complex contains an RNA polymerase promoter;   (b) adding RNA polymerase to form RNA products; and   (c) analyzing RNA products wherein an amount of RNA product is directly proportional to the amount of immunogen-binding agent-DNA-oligo complexes bound to the immobilized immunogens in step (a) and size of an RNA product corresponds to the step at which the immunogen-binding agent was added with earlier added immunogen-binding agents giving rise to longer products while later added immunogen-binding agents produce smaller products.   
     
     
         3 . A kit for detection and quantification of multiples immunogens in a sample via consecutive addition of quantifiable extenders comprising at least two epitope detectors of  claim 1 . 
     
     
         4 . A system for the detection and quantification of multiple immunogens in a sample, said system comprising a surface on which the epitope anchors are immobilized, and a plurality of epitope detectors of  claim 1 .