Application of cell surface-fluorescence immunosorbent assay in rapid screening of a hybridoma secreting monoclonal antibodyagainst PEDV N protein

ABSTRACT

To verify the effect of cell surface-fluorescence immunosorbent assay (CSFIA) on screening the highly active hybridoma cells, 6-8 week-old female BALB/c mice were immunized with recombinant porcine epidemic diarrhea virus (PEDV) N protein, and then spleen cells isolated from mice were fused with SP2/0 cells to generate hybridomas. The hybridomas secreting MAbs against PEDV N protein were screened by indirect ELISA and cell surface-fluorescence immunosorbent assay (CSFIA), respectively. The titer of MAbs secreted by each hybridoma was determined. Using these methods, two strains of hybridoma celllines secreting MAbs against PEDV N protein, designated as 3D3 and 1B1, respectively, were obtained and characterized. The results showed that the titer of 1B1 produced by CSFIA was 10 times higher than that of 3D3, and the preparation time was shortened by about 30 days on account of the direct selection of hybridoma through the fluorescence intensity without the need of multiple subcloning procedures. This study indicates that CSFIA is a powerful method to screen the hybridomas secreting higher titer MAb in a rapid one-step process, which would be valuable alternative for a rapid production of MAbs.