A toolbox of immunoprecipitation-grade monoclonal antibodies against human transcription factors

Anand Venkataraman, Kun Yang, Jose Irizarry, Mark Mackiewicz, Paolo Mita, Zheng Kuang, Lin Xue, Devlina Ghosh, Shuang Liu, Pedro Ramos, Shaohui Hu, Diane Bayron, Sarah Keegan, Richard Saul, Simona Colantonio, Hongyan Zhang, Florencia Pauli Behn, Guang Song, Edisa Albino, Lillyann AsencioLeonardo Ramos, Luvir Lugo, Gloriner Morell, Javier Rivera, Kimberly Ruiz, Ruth Almodovar, Luis Nazario, Keven Murphy, Ivan Vargas, Zully Ann Rivera-Pacheco, Christian Rosa, Moises Vargas, Jessica McDade, Brian S. Clark, Sooyeon Yoo, Seva G. Khambadkone, Jimmy de Melo, Milanka Stevanovic, Lizhi Jiang, Yana Li, Wendy Y. Yap, Brittany Jones, Atul Tandon, Elliot Campbell, Stephen Anderson, Richard M. Myers, Jef D. Boeke, David Fenyo, Gordon Whiteley, Joel S. Bader, Ignacio Pino, Daniel J. Eichinger, Heng Zhu, Seth Blackshaw

Research output: Contribution to journalArticlepeer-review

Abstract

A key component to overcoming the reproducibility crisis in biomedical research is the development of readily available, rigorously validated and renewable protein affinity reagents. As part of the NIH Protein Capture Reagents Program (PCRP), we have generated a collection of 1406 highly validated, immunoprecipitation (IP) and/or immunoblotting (IB) grade, mouse monoclonal antibodies (mAbs) to 736 human transcription factors. We used HuProt™ human protein microarrays to identify mAbs that recognize their cognate targets with exceptional specificity. Using an integrated production and validation pipeline, we validated these mAbs in multiple experimental applications, and have distributed them to the Developmental Studies Hybridoma Bank (DSHB) and several commercial suppliers. This study allowed us to perform a meta-analysis that identified critical variables that contribute to the generation of high quality mAbs. We find that using full-length antigens for immunization, in combination with HuProt™ analysis, provides the highest overall success rates. The efficiencies built into this pipeline ensure substantial cost savings compared to current standard practices.

Original languageEnglish (US)
JournalUnknown Journal
DOIs
StatePublished - Mar 14 2017
Externally publishedYes

Keywords

  • affinity
  • antibody specificity
  • ChIP-seq
  • effectiveness analysis
  • immunoprecipitation
  • mAb
  • monoclonal antibody
  • protein microarray

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Agricultural and Biological Sciences(all)
  • Immunology and Microbiology(all)
  • Neuroscience(all)
  • Pharmacology, Toxicology and Pharmaceutics(all)

Fingerprint Dive into the research topics of 'A toolbox of immunoprecipitation-grade monoclonal antibodies against human transcription factors'. Together they form a unique fingerprint.

Cite this