FOOD SCIENCE ›› 2010, Vol. 31 ›› Issue (13): 180-183.doi: 10.7506/spkx1002-6630-201013041

• Bioengineering • Previous Articles     Next Articles

Preparation and Identification of Anti-Lead-chelate Complex Polyclonal Antibody

ZHAI Yi-fan1,XIANG Jun-jian1,*,TANG Yong1,GUO Chang-wei1,LIU Xiao-feng2,YANG Hong-yu1   

  1. 1. Antibody Engineering Center, Jinan University, Guangzhou 510632, China ;
    2. Guangzhou Rivers Enterprise Co. Ltd., Guangzhou 510632, China
  • Received:2009-11-24 Online:2010-07-01 Published:2010-12-29
  • Contact: XIANG Jun-jian E-mail:txjj@jnu.edu.cn

Abstract:

Objective: BAS and OVA were individually conjugated with lead-chelate complex (Pb-DTPA) to prepare 2 antigens, namely Pb-DTPA-BSA, immunogen and Pb-DTPA-OVA, detective antigen. The synthesized antigens were then used to immunize BALB/c mice for preparing polyclonal antibodies. Finally, the antisera withdrawn from immunized BALB/c mice were assessed for their titers and specificities. Methods: The artificial antigens were identified by SDS-PAGE and scanned with ultraviolet spectrophotometer. The Pb2+ concentrations of the synthesized antigens were determined using inductively coupled plasma (ICP) spectrophotometer. Indirect ELISA and competitive ELISA were used to determine antiserum titer and specificity, respectively. Results: Compared with carrier proteins, the synthesized antigens exhibited lagged protein bands in SDS-PAGE pattern and blue shift of UV absorption peaks. The antiserum titers of the fourth and fifth mice reached up to 1:400,000 without cross-reactivity with OVA. The IC50 determined by IC-ELISA was 1.5 ng/mL and the cross reactivity with the other heavy ions were less than 1% although the cross-reactivity with Fe3+ was close to 5%. Conclusion: The above results suggest that anti-leadchelate complex polyclonal antibodies are prepared successfully.

Key words: lead ion, antigen synthesis, ELISA, polyclonal antibody

CLC Number: