Monoclonal Antibodies Against Male-Specific Antigen of White Lamphun Cattle (Bos indicus)

Abstract

A male-specific antibody was used in a cytotoxic reaction to increase X sperm in whole sperm by lysing Y sperm. The objective of this study was to produce a monoclonal antibody against male-specific antigen of White Lamphun cattle. From this study, monoclonal antibodies were produced by biweekly immunization of 3 Balb/c mice with 10 × 106 cells of White Lamphun cattle sperm. Mouse number 2 was chosen to be used in the fusion process by fusing myeloma cells and splenocytes from the immunized Balb/c female mouse with a ratio of 3:1 (splenocyte:myeloma) and the result from the enzyme-linked immunosorbent assay (ELISA) showed that hybridoma cells named P1C2 produced antibody amounts against male and female white blood cells of White Lamphun cattle of 0.55 and 0.38 optical density (O.D.) units. P1C2 was used in the cell limiting process, evaluated using antibody titer and resulted in positive clones named P1C2B8 and P1C2C9. The ELISA results showed that P1C2B8, P1C2C9 and P1C2B8&C9 produced antibody amounts against male and female white blood cells of 1.74 ± 0.03 and 1.56 ± 0.05, 1.04 ± 0.03 and 0.88 ± 0.03 and 1.47 ± 0.07 and 1.27 ± 0.07 O.D. units, respectively. The results from immunofluorescence showed that P1C2B8, P1C2C9 and P1C2B8&C9 produced antibody amounts against male and female white blood cells of 92.15 ± 0.87 and 69.33 ± 0.93, 91.19 ± 0.95 and 72.95 ± 1.27 and 93.27 ± 1.25 and 68.68 ± 1.23 %, respectively (P<0.05). It was concluded that the monoclonal antibodies produced in this study were specific to the male more than the female.