Development of Monoclonal Hybridoma Cell Lines and Extracting Antibody Against Fummonisin B₁
Pages 507-513
Guangyu Yang, Wenxue Li, Xiaoyun Liang, Jinyin Wu and Wei Zhu

DOI: http://dx.doi.org/10.6000/1927-5129.2015.11.68

Published: 10 September 2015

Abstract: Objective:To acquire monoclonal hybridoma cell lines against fummonisin B₁(FB₁) and extract monoclonal antibody against FB₁.

Methods: Coupling antigens of FB₁-KLH and FB₁-BSA with chemical methods and immune 6-8 weeks old female BALB/c mice with FB₁-KLH. Integrating spleen cells with sp2/0 myeloma cells to acquire hybridoma cell lines secreting McAb against FB₁. The method of multiple subclones was used to select cell lines stably secreting McAb. McAbs was got from ascites and purified. The subclass of antibody was measured and the molecular weight was identified. The specificity and sensitivity of McAb were identified with indirect competitive inhibition ELISA.

Results:The results of serum from immuned mice showed that after five times of immunization the titer stables at 1×10^-6, and the McAb belongs to IgG1 subclass, the light chain was κ, the molecular weight of heavy and light chain were 55kDa and 32kDa, respectively. ELISA results showed that McAb could react with FB₁. The linear range indirect competitive inhibition ELISA is 10-500ng/ml.

Conclusion:The monoclonal hybridoma cell lines and the high specificity,high sensitivity of FB1-McAb was
got.

Keywords: Fummonisin B₁, hybridomacell lines, monoclonal antibody.