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Generation of Murine Monoclonal Antibodies by Hybridoma Technology
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A Rapid Method for Antigen-Specific Hybridoma Clone Isolation.

Xiuqing Li1, Hongfen Bian1, Siming Yu2

  • 1Department of Bioengineering, Guangdong Province Engineering Research Center for Antibody Drug and Immunoassay, Jinan University , Guangzhou 510632, P. R. China.

Analytical Chemistry
|January 2, 2018
PubMed
Summary
This summary is machine-generated.

A new cell surface fluorescence immunosorbent assay (CSFIA) rapidly isolates antigen-specific hybridoma clones. This efficient method significantly reduces processing time and labor compared to traditional techniques for monoclonal antibody production.

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Area of Science:

  • Biotechnology
  • Immunology
  • Cell Biology

Background:

  • Traditional methods for isolating antigen-specific hybridoma clones, such as ELISA and limited dilution, are time-consuming and labor-intensive.
  • Efficient isolation of high-quality hybridoma clones is crucial for monoclonal antibody production.

Purpose of the Study:

  • To develop and validate a simple, rapid, and efficient cell surface fluorescence immunosorbent assay (CSFIA) for detecting and isolating antigen-specific hybridoma clones.
  • To compare the efficiency and speed of the CSFIA method with conventional ELISA-based approaches.

Main Methods:

  • Antigen anchoring to hybridoma cell surfaces using Oleyl-PEG4000-NHS.
  • Detection of secreted antibodies via cell-surface-bound antigens.
  • Fluorescent staining of positive hybridoma cells using anti-mouse IgG-Fc antibody.
  • Isolation of positive clones from methylcellulose semisolid medium.

Main Results:

  • The CSFIA method successfully selected antigen-specific hybridoma clones against malachite green and porcine epidemic diarrhea virus with high efficiency.
  • CSFIA isolated over two times more hybridoma clones compared to ELISA-based methods.
  • The CSFIA method required less than 25% of the processing time of ELISA-based methods.

Conclusions:

  • The CSFIA is a highly efficient, inexpensive, and simple method for isolating antigen-specific hybridoma clones.
  • CSFIA offers a significant improvement in speed and yield over traditional methods for monoclonal antibody preparation.
  • This assay is an excellent candidate for routine use in hybridoma screening and monoclonal antibody development.