In order to evaluate the effect of heavy metal artificial antigen using heterocyclic Bifunctional Chelators（BFCs），S-2-（4-Aminobenzyl）-1，4，7，10-tetraazacyclododecane tetraacetic acid（P-NH2-Bn-DOTA， eg. DOTA）and heavy metal Pb2+ were selected. The artificial antigens were produced by glutaraldehyde method based on the Bifunctional Chelators DOTA. The conjugation ratios were 13 ∶ 1 and 9 ∶ 1 for immunizing antigen and coating antigen， respectively. The polyclonal antibody（PcAb） was obtained after immuring to rabbits. Based on the PcAb， an indirect competitive ELISA （ic-ELISA） was established and its linear regression equation was y=12.014ln（x）-13.249 （R2=0.9951）. According to the ic-ELISA， the value of IC50 and IC20 were 193.4 ng/mL and 15.9 ng/mL， respectively. The specificity analysis showed that the cross-reactivities （CR） were all less 6.4% except Cd2+ was 18%， which indicated that the PcAb was specific for Pb2+. The produced Cu2+ artificial antigen and the PcAb could lay the foundation for rapid analysis of heavy metal residue using the BFCs.