Journal article
A Rapid Method for Antigen-Specific Hybridoma Clone Isolation
Analytical chemistry (Washington), Vol.90(3), pp.2224-2229
02/06/2018
PMID: 29290124
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.
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Details
- Title
- A Rapid Method for Antigen-Specific Hybridoma Clone Isolation
- Creators
- Xiuqing Li - Department of Bioengineering, Guangdong Province Engineering Research Center for Antibody Drug and Immunoassay, Jinan University, Guangzhou 510632, P. R. ChinaHongfen Bian - Jinan UniversitySiming Yu - Jinan UniversityWei Xiao - Jinan UniversityJianying Shen - Guangzhou Highway Engineering Company, Guangzhou 510075, P. R. ChinaCaifeng Lan - Jinan UniversityKenan Zhou - Jinan UniversityCaihong Huang - Jinan UniversityLei Wang - Jinan UniversityDan Du - Washington State UniversityYuehe Lin - Washington State UniversityYong Tang - Jinan University
- Publication Details
- Analytical chemistry (Washington), Vol.90(3), pp.2224-2229
- Academic Unit
- School of Mechanical and Materials Engineering
- Number of pages
- 6
- Identifiers
- 99901227851401842
- Language
- English
- Resource Type
- Journal article