[Preparation and identification of monoclonal antibodies against Helicobacter pylori].
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 (K(aff)) ranging from 1 x 10(-10) to 5.2 x 10(-12) mol/L.


CONCLUSIONS
The mAbs specially against Hp have been obtained, which may facilitate further study of detection and vaccine development of Hp.
Li
Yan
Y
School of Biotechnology, Southern Medical University, Guangzhou 510515, China. lyinf@ 163.com
Ning
Yun-shan
YS
Hong
Yan-hua
YH
Liu
Yi-chu
YC
Luo
Jun
J
Long
Min
M
Dong
Wen-qi
WQ
Li
Ming
M
D000818:Animals; D000907:Antibodies, Bacterial / Q000096:biosynthesis* / Q000276:immunology; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D000918:Antibody Specificity; D005260:Female; D016480:Helicobacter pylori / Q000276:immunology*; D006801:Humans; D006825:Hybridomas / Q000276:immunology; D051379:Mice; D008807:Mice, Inbred BALB C
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal