Preparation and identification of monoclonal antibodies against Helicobacter pylori / 南方医科大学学报

ABSTRACT

<p><b>OBJECTIVE</b>To prepare and identify monoclonal antibodies (mAbs) against Helicobacter pylori (Hp).</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>The mAbs specially against Hp have been obtained, which may facilitate further study of detection and vaccine development of Hp.</p>