Objective: To establish hybridoma cell lines against sporozoites of Eimeria acervulina.
Methods: BALB/c mice were immunized with purified sporozoites of E. acervulina. Hybridoma cell lines were set up by using hybridoma technique, and monoclonal antibodies were prepared. The monoclonal antibody was identified by determining their cross reactivity, relative affinity, immunoglobulin class or subclass with enzyme linked immunosorbent assay (ELISA).
Results: Four hybridoma cell lines stably secreting McAbs against sporozoites were obtained: Easp-3G3 and Easp-5G10 belonging to IgG1, Easp-3H6 belonging to IgG2b, Easp-5H4 belonging to IgG2a. All four McAbs bound with E. acervulina sporozoite protein, but the Easp-5H4 McAb showed cross reactivity with E. tenellum sporozoite protein. Different antigenic epitopes were recognized by Easp-3G3 or Easp-5G10 and Easp-3H6 or Easp-5H4.
Conclusion: Three of the four produced monoclonal antibodies show high specificity and affinity to the Eimeria acervulina sporozoites.