Rapid production of antigen-specific monoclonal antibodies from a variety of animals

BMC Biology

Methodology Article

Rika Fujimoto¹  Fuminori Yamagishi²  Megumi Yoshioka³  Masaharu Isobe⁴  Nobuyuki Kurosawa⁴ 

[1] Department of Immunology, Kochi Medical School, Nangoku-shi, 783-8505, Kochi, Japan;Department of Surgery, Itoigawa General Hospital, Itoigawa-shi, 941-0006, Niigata, Japan;Graduate School of Innovative Life Science, University of Toyama, Toyama-shi, 930-8555, Toyama, Japan;Laboratory of Molecular and Cellular Biology, Faculty of Science and Engineering, Graduate School, University of Toyama, 3190 Gofuku, Toyama-shi, 930-8555, Toyama, Japan;

关键词: antigen-specific monoclonal antibody;    ER-tracker;    ERIAA;    FACS;    guinea pig;    human;    MAGrahd;    rabbit;    rat;    single cell;    TS-jPCR;   

DOI  :  10.1186/1741-7007-10-80

 received in 2012-07-10, accepted in 2012-09-28,  发布年份 2012

来源: Springer

【 摘 要 】

BackgroundAlthough a variety of animals have been used to produce polyclonal antibodies against antigens, the production of antigen-specific monoclonal antibodies from animals remains challenging.ResultsWe propose a simple and rapid strategy to produce monoclonal antibodies from a variety of animals. By staining lymph node cells with an antibody against immunoglobulin and a fluorescent dye specific for the endoplasmic reticulum, plasma/plasmablast cells were identified without using a series of antibodies against lineage markers. By using a fluorescently labeled antigen as a tag for a complementary cell surface immunoglobulin, antigen-specific plasma/plasmablast cells were sorted from the rest of the cell population by fluorescence-activated cell sorting. Amplification of cognate pairs of immunoglobulin heavy and light chain genes followed by DNA transfection into 293FT cells resulted in the highly efficient production of antigen-specific monoclonal antibodies from a variety of immunized animals.ConclusionsOur technology eliminates the need for both cell propagation and screening processes, offering a significant advantage over hybridoma and display strategies.

【 授权许可】

Unknown   
© Kurosawa et al; licensee BioMed Central Ltd. 2012. This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.