Rapid identification of monospecific monoclonal antibodies using a human proteome microarray.

Published

Journal Article

To broaden the range of tools available for proteomic research, we generated a library of 16,368 unique full-length human ORFs that are expressible as N-terminal GST-His(6) fusion proteins. Following expression in yeast, these proteins were then individually purified and used to construct a human proteome microarray. To demonstrate the usefulness of this reagent, we developed a streamlined strategy for the production of monospecific monoclonal antibodies that used immunization with live human cells and microarray-based analysis of antibody specificity as its central components. We showed that microarray-based analysis of antibody specificity can be performed efficiently using a two-dimensional pooling strategy. We also demonstrated that our immunization and selection strategies result in a large fraction of monospecific monoclonal antibodies that are both immunoblot and immunoprecipitation grade. Our data indicate that the pipeline provides a robust platform for the generation of monoclonal antibodies of exceptional specificity.

Full Text

Duke Authors

Cited Authors

  • Jeong, JS; Jiang, L; Albino, E; Marrero, J; Rho, HS; Hu, J; Hu, S; Vera, C; Bayron-Poueymiroy, D; Rivera-Pacheco, ZA; Ramos, L; Torres-Castro, C; Qian, J; Bonaventura, J; Boeke, JD; Yap, WY; Pino, I; Eichinger, DJ; Zhu, H; Blackshaw, S

Published Date

  • June 2012

Published In

Volume / Issue

  • 11 / 6

Start / End Page

  • O111.016253 -

PubMed ID

  • 22307071

Pubmed Central ID

  • 22307071

Electronic International Standard Serial Number (EISSN)

  • 1535-9484

International Standard Serial Number (ISSN)

  • 1535-9476

Digital Object Identifier (DOI)

  • 10.1074/mcp.O111.016253

Language

  • eng