Abstract: In order to produce monoclonal antibodies (McAbs) against bursin, the artificial antigens of bursin (BSA-KHG-NH₂and OVA-KHG-NH₂) were prepared by EDC method, in which the amino group on -Lys of bursin molecules was conjugated to bovine serum albumin (BSA) or chicken ovalbumin (OVA), respectively. BALB/c mice were immunized four times by BSA-KHG-NH₂, and eighteen hybridoms producing anti-bursin monoclonal antibodies were obtained by limited dilution. The binding of artificial antigen OVA-KHG-NH₂ to the McAbs were competitively inhibited by free bursin(KHG-NH₂) and GKHG-NH₂peptide,respectively, but couldn't be inhibited by free KHGK peptide. These results indicated that the 18 McAbs specifically bonded to bursin molecules. An IC-ELISA method to detect bursin was developed using McAb 2D2, in which the detection limits was about 4 ng/mL and the linear range of the inhibition curve was about 4 to 500 ng/mL, the recovery rate of bursin in the samples ranged from 85.0% to 92.4%.

Key words: bursin; artificial antigen; monoclonal antibodies; IC-ELISA