“A key component of efforts to address the reproducibility crisis in biomedical research is the development of rigorously validated and renewable protein-affinity reagents. As part of the US National Institutes of Health (NIH) Protein Capture Reagents Program (PCRP), we have generated a collection of 1,406 highly validated immunoprecipitation- and/or immunoblotting-grade mouse monoclonal antibodies (mAbs) to 737 human transcription factors, using an integrated production and validation pipeline. We used HuProt human protein microarrays as a primary validation tool to identify mAbs with high specificity for their cognate targets. We further validated PCRP mAbs by means of multiple experimental applications, including immunoprecipitation, immunoblotting, chromatin immunoprecipitation followed by sequencing (ChIP-seq), and immunohistochemistry. We also conducted a meta-analysis that identified critical variables that contribute to the generation of high-quality mAbs.

All validation data, protocols, and links to PCRP mAb suppliers are available at http://proteincapture.org.”

Pipeline description
Secondary validation of PCRP reagents.
The attrition funnel through which immunogens entering the PCRP pipeline ultimately generate high-quality mAbs.

VenkataramanA, YangK, IrizarryJ, Mackiewicz M, MitaP, KuangK XueL, Ghosh D, LiuS, RamosP, Hu S, Bayron D, Keegan S, Saul R, Colantonio S, Zhang H, Behn FP, Song G, Albino E, Asencio L, Ramos L, Lugo L, Morell G, Rivera J, Ruiz K, Almodovar R, Nazario L, Murphy K, Vargas I, Rivera-Pacheco ZA, Rosa C, Vargas M, McDade J, Clark BS, Yoo S, Seva G. Khambadkone5, de Melo J, Stevanovic M, Jiang L, Li Y, Yap WY, Jones B, Tandon A, CampbellE, AndersonS, Myers RM, BoekeJD, FenyoD,  Whiteley G, BaderJS, Pino I, Eichinger DJ, ZhuH, and BlackshawS.  A toolbox of immunoprecipitation-grade monoclonal antibodies against human transcription factors. bioRxiv,  017, 116442; doi: https://doi.org/10.1101/116442.