A protein microarray ELISA for screening biological fluids.
Protein microarrays permit the simultaneous measurement of many proteins in a small sample volume and therefore provide an attractive approach for the quantitative measurement of proteins in biological fluids, including serum. This chapter describes a microarray enzyme-linked immunosorbent assay (ELISA). Capture antibodies are immobilized onto a glass surface; the covalently attached antibodies bind a specific antigen from a sample overlaying the array. A second, biotinylated antibody that recognizes the same antigen as the first antibody, but at a different epitope, is then used for detection. Detection is based on an enzymatic signal-enhancement method known as tyramide signal amplification (TSA). By coupling a microarray-ELISA format with the signal amplification of tyramide deposition, the assay sensitivity is as low as sub-pg/mL.
Varnum
Susan M
SM
Biological Sciences Division, Pacific Northwest National Laboratory, Richland, WA, USA.
Woodbury
Ronald L
RL
Zangar
Richard C
RC
D000577:Amides / Q000737:chemistry / Q000378:metabolism; D000906:Antibodies / Q000378:metabolism; D000915:Antibody Affinity; D001710:Biotin / Q000378:metabolism; D001826:Body Fluids / Q000737:chemistry*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D000939:Epitopes; D040081:Protein Array Analysis / Q000379:methods*; D011506:Proteins / Q000032:analysis*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D019809:Streptavidin / Q000378:metabolism
D000577:Amides; D000906:Antibodies; D000939:Epitopes; D011506:Proteins; D001710:Biotin; D019809:Streptavidin