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Open Access Highly Accessed Methodology article

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

Nobuyuki Kurosawa1, Megumi Yoshioka2, Rika Fujimoto3, Fuminori Yamagishi4 and Masaharu Isobe1*

Author Affiliations

1 Laboratory of Molecular and Cellular Biology, Faculty of Science and Engineering, Graduate School, University of Toyama, 3190 Gofuku, Toyama-shi, Toyama, 930-8555, Japan

2 Graduate School of Innovative Life Science, University of Toyama, Toyama-shi, Toyama, 930-8555, Japan

3 Department of Immunology, Kochi Medical School, Nangoku-shi, Kochi, 783-8505, Japan

4 Department of Surgery, Itoigawa General Hospital, Itoigawa-shi, Niigata, 941-0006, Japan

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BMC Biology 2012, 10:80  doi:10.1186/1741-7007-10-80

Published: 28 September 2012

Abstract

Background

Although a variety of animals have been used to produce polyclonal antibodies against antigens, the production of antigen-specific monoclonal antibodies from animals remains challenging.

Results

We 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.

Conclusions

Our technology eliminates the need for both cell propagation and screening processes, offering a significant advantage over hybridoma and display strategies.

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