Multiplexed protein detection by proximity ligation for cancer biomarker validation.
We present a proximity ligation-based multiplexed protein detection procedure in which several selected proteins can be detected via unique nucleic-acid identifiers and subsequently quantified by real-time PCR. The assay requires a 1-microl sample, has low-femtomolar sensitivity as well as five-log linear range and allows for modular multiplexing without cross-reactivity. The procedure can use a single polyclonal antibody batch for each target protein, simplifying affinity-reagent creation for new biomarker candidates.
Fredriksson
Simon
S
Stanford Genome Technology Center, Stanford, California 94305, USA. simon.fredriksson@stanford.edu
Dixon
William
W
Ji
Hanlee
H
Koong
Albert C
AC
Mindrinos
Michael
M
Davis
Ronald W
RW
D000906:Antibodies / Q000032:analysis*; D014408:Biomarkers, Tumor / Q000032:analysis*; D009363:Neoplasm Proteins / Q000032:analysis*; D009369:Neoplasms / Q000175:diagnosis* / Q000378:metabolism; D009841:Oligonucleotides / Q000032:analysis; D010865:Pilot Projects; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D000906:Antibodies; D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D009841:Oligonucleotides