Proteomics-based generation and characterization of monoclonal antibodies against human liver mitochondrial proteins.

Published

Conference Paper

Monoclonal antibodies (mAbs) have the potential to be a very powerful tool in proteomics research to determine protein expression, quantification, localization and modification, as well as protein-protein interactions, especially when combined with microarray technology. Thus, a large amount of well-characterized and highly qualified antibodies are needed in proteomics. Purified antigen, which is not always available, has proven to be one of the rate-limiting steps in mAb large-scale generation. Here we describe our strategies to establish a murine hybridoma cell bank for human liver mitochondria using unknown native proteins as the immunogens. The antibody-recognized mitochondrial proteins were identified by MS following immunoprecipitation (IP), and by screening of human liver cDNA expression library. We found that the established antibodies reacted specifically with a number of important enzymes in mitochondria. The subcellular localization of these antigens in mitochondria was further confirmed by immunohistocytochemistry. A panel of antibodies was also tested for their ability to capture and deplete the targeting proteins and complexes from the total mitochondrial proteins. We believe these well-characterized antibodies would be useful in various applications for Human Liver Proteome Project (HLPP) when the scale of this hybridoma cell bank is enlarged significantly in the near future.

Full Text

Cited Authors

  • Gao, J; Gao, Y; Ju, Y; Yang, J; Wu, Q; Zhang, J; Du, X; Wang, Z; Song, Y; Li, H; Luo, X; Ren, F; Li, J; Chen, Y; Wang, L; Xu, H; Liu, X; Wang, J; Zhang, Y; Cai, Y; Cui, Y; Qian, X; He, F; Li, M; Sun, Q-H

Published Date

  • January 2006

Published In

Volume / Issue

  • 6 / 2

Start / End Page

  • 427 - 437

PubMed ID

  • 16342244

Pubmed Central ID

  • 16342244

Electronic International Standard Serial Number (EISSN)

  • 1615-9861

International Standard Serial Number (ISSN)

  • 1615-9853

Digital Object Identifier (DOI)

  • 10.1002/pmic.200500409