Preparation and identification of monoclonal antibodies against Helicbacter pylori

Abstract

Abstract: Objective To prepare and identify monoclonal antibodies (mAbs) against Helicobacter pylori (Hp). Methods BALB/c mice were immunized with the supernatant and precipitation of cultured Hp after ultrasonication and mAbs were obtained by means of hybridoma technique. The resultant mAbs was evaluated for subtype, titer, affinity, and further identified with Lpp20, HspA, urease A, CagA, urease B, and catalase prepared by recombinant expression. Results Totally 34 hybridoma cell lines were established which secreted specific mAbs, including 31 against the supernatant and 3 against the precipitation of Hp, and the prepared mAbs showed specific reaction against Lpp20 (3 strains), HspA (2 strains), urease A (4 strains), CagA (1 strain), urease B (5 strains), and catalase (2 strains) antigens, respectively. The mAbs was all identified as immunoglobulin G1 (IgG1) and theirs titer in the culture supernatant and ascites was 1:16 to 1:32 and 1: 32000 to 1:64000 respectively with affinity constants (Kaff) ranging from 1×10-10 to 5.2×10-12 mol/L. Conclusion The mAbs specially against Hp have been obtained, which may facilitate further study of detection and vaccine development of Hp.

CLC Number:

R392

LI Yan, NING Yun-shan, HONG Yan-hua, LIU Yi-chu, LUO Jun, LONG Min, DONG Wen-qi, LI Ming School of Biotechnology, Department of Microbiology, School of Public Health and Tropical Disease, Southern Medical University, Guangzhou 510515, China. Preparation and identification of monoclonal antibodies against Helicbacter pylori[J]. Journal of Southern Medical University, 2006, 26(04): 425-427.

References

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