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Anal Chem. 2018 Feb 6;90(3):2224-2229. doi: 10.1021/acs.analchem.7b04595. Epub 2018 Jan 17.

A Rapid Method for Antigen-Specific Hybridoma Clone Isolation.

Author information

1
Department of Bioengineering, Guangdong Province Engineering Research Center for Antibody Drug and Immunoassay, Jinan University , Guangzhou 510632, P. R. China.
2
Key Laboratory of Biomaterials of Guangdong Higher Education Institutes, Department of Biomedical Engineering, Jinan University , Guangzhou 510632, China.
3
Guangzhou Highway Engineering Company , Guangzhou 510075, P. R. China.
4
School of Mechanical and Materials Engineering, Washington State University , Pullman, Washington 99164, United States.
5
Institute of Food Safety and Nutrition, Jinan University , Guangzhou 510632, China.

Abstract

Using an enzyme-linked immunosorbent assay (ELISA) and limited dilution methods to screen and clone antigen-specific hybridoma cells is extremely time-consuming and labor-intensive. This work features a simple and rapid cell surface fluorescence immunosorbent assay (CSFIA), designed for the detection and isolation of antigen-specific hybridoma clones. In this assay, antigens are first anchored to the hybridoma cell surface through a dual-functioning molecular Oleyl-PEG4000-NHS. Specific antibodies secreted from hybridoma cells are then captured by the antigens on the cell surface. Positive hybridoma cells are stained using a fluorescently labeled anti-mouse IgG-Fc antibody. After the addition of a methylcellulose semisolid medium, positive clones are easily picked using a pipet. These positive cell clones can be used to produce monoclonal antibodies after direct expansion. Using this method, positive hybridoma clones against both malachite green and porcine epidemic diarrhea virus are selected with high efficiency. Compared to the ELISA-based method, the CSFIA-based method achieved the capability of isolating >2-fold more hybridoma clones in <25% of the corresponding processing time. In brief, the CSFIA-based method is highly efficient and inexpensive with a simple and direct operation, which is an excellent candidate method for antigen-specific positive clone isolation in a monoclonal antibody preparation.

PMID:
29290124
DOI:
10.1021/acs.analchem.7b04595
[Indexed for MEDLINE]

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