Synthesis and Identification of SG-BSA and SG-OVA

Hapten sulfaguanidine (SG) was coupled with carrier protein bovine serum albumin (BSA) to form a full antigen SG-BSA by diazotization and glutaraldehyde methods. Ovalbumin (OVA) was used as protein carrier to couple with Hapten SG by glutaraldehyde method. Then, the immunogen and the coating antigen were purified by dialysis and gel exclusion chromatography. The conjugated ratio of SG to BSA in artificial antigen was 5.3 (using diazotization method) and 6.5 (glutaraldehyde method), and the conjugated ratio of SG to OVA in coating antigen was 2.3 (glutaraldehyde method) by UV-visible spectrophotometer. The coupling was successful according to the analysis of sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE). BALB/c mice were immunized with the antigen (SG-BSA), and the titers of antiserum were tested to be 1 : 6 400 and 1 : 400 after three periods of immunities by indirect ELISA, which further identified the success of the synthesis of both immunogen SG-BSA and coating antigen SG-OVA.