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A Rapid Method for Antigen-Specific Hybridoma Clone Isolation

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 e...

Authors:   Xiuqing Li; Hongfen Bian; Siming Yu; Wei Xiao; Jianying Shen; Caifeng Lan; Kenan Zhou; Caihong Huang; Lei Wang; Dan Du; Yuehe Lin; Yong Tang
Journal:   Analytical Chemistry
Year:   2018
DOI:   10.1021/acs.analchem.7b04595
Publication date:   17-Jan-2018
Facts, background information, dossiers
  • hybridoma
  • monoclonal antibodies
  • Malachite green
  • diarrhea
  • antigens
  • antibodies
More about American Chemical Society Publications
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