Objective: To compare the immunogenity of the TTX artificial antigen with two different carrier proteins, bovine serum albumin (BSA) and keyhole limpet hemocyanin (KLH), and antiserum against each immunogen were well investigated.
Methods: The hapten Tetrodotoxins (TTX) were linked to carrier protein BSA or KLH as immunogen, and linked ovalbumin (OVA) as coating antigen by the Mannich method. Then the 6-8 weeks Babl/c mice were immunized with TTX-BSA or TTX-KLH. And the TTX-BSA also was another coating antigen in the detection of TTX-KLH antiserum. The antiserum was detected by indirect enzyme-linked immunosorbent assay (iELISA) and indirect competitive enzyme-linked immunosorbent assay (icELISA).
Results: The high titer of antiserum was 1:64000 after immunizing mouse. In the detection of TTX-KLH antiserum, the A450 value with the TTX-OVA coating antigens was a little higher than the TTX-BSA coating antigen, and IC50 was 10-20 ng/ml by icELISA.
Conclusion: The artificial antigen was successfully synthesized, and the conjugate of TTX-KLH was testified to have a better immunogenity in comparison with TTX-BSA, this investigation will benefit development of TTX immunoassay.