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

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	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
Description	Country affiliation: United States Author Affiliation: Jeong JS ( Department of Pharmacology, Johns Hopkins University School of Medicine, Baltimore, Maryland, USA.)
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.
File Format	HTM / HTML
ISSN	15359476
e-ISSN	15359484
DOI	10.1074/mcp.O111.016253
Journal	Molecular & Cellular Proteomics
Issue Number	6
Volume Number	11
Language	English
Publisher	American Society for Biochemistry and Molecular Biology
Publisher Date	2012-06-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Proteomics Antibodies, Monoclonal, Murine-derived Immunology Antibody Specificity Proteome Recombinant Fusion Proteins Animals Antigens Chemistry Cell Line, Tumor Hybridomas Immobilized Proteins Mice Mice, Inbred Balb C Protein Array Analysis Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Medicine Analytical Chemistry Molecular Biology Biochemistry

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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