The NIH Protein Capture Reagents Program (PCRP): a standardized protein affinity reagent toolbox

• Published in: Nature methods Vol. 13; no. 10; pp. 805 - 806
• Main Authors: Blackshaw, Seth, Venkataraman, Anand, Irizarry, Jose, Yang, Kun, Anderson, Stephen, Campbell, Elliot, Gatlin, Christine L, Freeman, Nancy L, Basavappa, Ravi, Stewart, Randall, Loss, Michael A, Pino, Ignacio, Zhu, Heng, Bader, Joel S
• Format: Journal Article
• Language: English
• Published: United States Nature Publishing Group 01.10.2016
• Subjects: Antibodies; Antibodies, Monoclonal - immunology; Antigens; Chromatography, Affinity - standards; Genetic aspects; Humans; Identification and classification; Immunoassay - methods; Indicators and Reagents - chemistry; Innovations; National Institutes of Health (U.S.); Peptide Library; Pilot projects; Proteins; Proteins - analysis; Proteins - chemistry; Proteins - immunology; Proteomics; United States
• ISSN: 1548-7091; 1548-7105
• DOI: 10.1038/nmeth.4013

In 2010, the National Institutes of Health launched a pilot program to generate and distribute high-quality affinity reagents against human transcription factors (hTFs) and transcriptional coregulators. The NIH Common Fund Protein Capture Reagents Program (PCRP) production pipeline consists of three core centers: antigen production, traditional monoclonal antibody (mAb) production, and recombinant phage displayed antibody binding fragment (rAb) production. The PCRP differs in important ways from previous efforts to generate collections of affinity reagents for human proteins3,4. First, mAbs and rAbs are generated, and these molecules allow for greater ease of characterization and reproducibility than do polyclonal antibodies. Second, individual protein domains and full-length proteins are used as antigens, enriching for rAbs and mAbs that recognize proteins in native conformation. Third, each mAb or rAb is subjected to a high-throughput initial screen of either affinity or specificity; each mAb or rAb is only sent for further validation if it passes this screen. Fourth, each reagent that passes this primary screen is subject to multiple types of secondary validation, including immunoprecipitation (IP) and western blotting (WB). Finally, all reagents are made available to the community at low cost through commercial and not-for-profit sources.