[Preparation and selection of the monoclonal antibody used for kits to detect BNP32 which was applied to clinical research].

AIM

To prepare the monoclonal antibody (mAb) used for kits to detect the BNP32 antigen by means of double-antibody sandwich ELISA assay. Comparasion differences on detection BNP between double-antibody sandwich ELISA and Roche ECL.

METHODS

BALB/c mice were immunized with purified recombinant BNP32 protein and by routine hybridoma technique, Then comparasion differences on detection BNP between double-antibody sandwich ELISA and Roche ECL.

RESULTS

16 hybridoma cell lines secreting potent mAb against BNP32 were obtained. The subtype of these 16 mAb were found to be IgG1, IgG2a and IgM, then their cross-blocking properties were analyzed, when they reacted with the BNP32 protein in direct ELISA in order to offer the valuable data for selecting feasible pair of mAb in the detection of the BNP32 antigen. All the mAbs were used for the detection of BNP32 by double-antibody sandwich ELISA. In addition, the mAbs were purified and HRP-labelled in advance. Finally, we had successfully screened a pair of mAbs, which exhibited a sensitivity of 20 ng/L for the detection of BNP32 antigen. The two detection methods have very good consistency (kappa=0.828>0.75)between double-antibody sandwich ELISA and Roche ECL. There was no statistics significance on differences between these two methods(P>0.05).

CONCLUSION

It is evident that this pair of mAb shows excellent detection of BNP32. The double-antibody sandwich ELISA can be good apply to detect BNP level in clinical congestive heart failure patients.