Preparation and detection of antisera for collagen protein typeII

Objective: To investigate the regular patterns of antibody production and prepare the antiserum on collagen protein typeⅡ(CⅡ).Methods: The New Zealand white rabbits were immunized with purified chicken and pig CⅡ and then the rabbit sera were collected every 10 days.The improved indirect ELISA method was adopted to detect the titers of anti-CⅡantibodies. Results: When the rabbits were immunized with purified chicken CⅡ,the antibody titers in sera were 1∶3 200 at 10 th day and 1∶204 800 at 20th day.After booster the antibody titers increased to 1∶409 600 at 30 th day and 1∶819 200 at 40 th day.When the rabbits were immunized with purified pig CⅡ,the antibodies could not be detected at 10 th day.But at 20 th day the antibody titer was 1∶3 200.After booster the antibody titers were then increased to 1∶51 200 at 30 th day and 1∶102 400 at 40 th day. Conclusion: The anti-CⅡ antibodies can emerge in serum after latency and the antiserum with high titer can be obtained when the rabbits were immunized with purified chicken and pig CⅡ.The antibody production was consistent with relevant regularity of primary and secondary immune responses.The latent phases and titers of anti-CⅡ antibodies in sera were concerned with collagen proteins from different genus.