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

Jun Seop Jeong, Lizhi Jiang, Edisa Albino, Josean Marrero, Hee Sool Rho, Jianfei Hu, Shaohui Hu, Carlos Vera, Diane Bayron-Poueymiroy, Zully Ann Rivera-Pacheco, Leonardo Ramos, Cecil Torres-Castro, Jiang Qian, Joseph Bonaventura, Jef D. Boeke, Wendy Y. Yap, Ignacio Pino, Daniel J. Eichinger, Heng Zhu, Seth Blackshaw

Research output: Contribution to journalArticle

Abstract

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.

Original languageEnglish (US)
JournalMolecular and Cellular Proteomics
Volume11
Issue number6
DOIs
StatePublished - Jun 2012

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Antibody Specificity
Proteome
Microarrays
Immunization
His-His-His-His-His-His
Monoclonal Antibodies
Tissue Array Analysis
Fungal Proteins
Antibodies
Microarray Analysis
Immunoprecipitation
Proteomics
Open Reading Frames
Libraries
Fusion reactions
Pipelines
Cells
Research
Proteins

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Rapid identification of monospecific monoclonal antibodies using a human proteome microarray. / Jeong, Jun Seop; Jiang, Lizhi; Albino, Edisa; Marrero, Josean; Rho, Hee Sool; Hu, Jianfei; Hu, Shaohui; Vera, Carlos; Bayron-Poueymiroy, Diane; Rivera-Pacheco, Zully Ann; Ramos, Leonardo; Torres-Castro, Cecil; Qian, Jiang; Bonaventura, Joseph; Boeke, Jef D.; Yap, Wendy Y.; Pino, Ignacio; Eichinger, Daniel J.; Zhu, Heng; Blackshaw, Seth.

In: Molecular and Cellular Proteomics, Vol. 11, No. 6, 06.2012.

Research output: Contribution to journalArticle

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 2012, 'Rapid identification of monospecific monoclonal antibodies using a human proteome microarray.', Molecular and Cellular Proteomics, vol. 11, no. 6. https://doi.org/10.1074/mcp.O111.016253
Jeong, Jun Seop ; Jiang, Lizhi ; Albino, Edisa ; Marrero, Josean ; Rho, Hee Sool ; Hu, Jianfei ; Hu, Shaohui ; Vera, Carlos ; Bayron-Poueymiroy, Diane ; Rivera-Pacheco, Zully Ann ; Ramos, Leonardo ; Torres-Castro, Cecil ; Qian, Jiang ; Bonaventura, Joseph ; Boeke, Jef D. ; Yap, Wendy Y. ; Pino, Ignacio ; Eichinger, Daniel J. ; Zhu, Heng ; Blackshaw, Seth. / Rapid identification of monospecific monoclonal antibodies using a human proteome microarray. In: Molecular and Cellular Proteomics. 2012 ; Vol. 11, No. 6.
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