[Development of panel of monoclonal antibodies for detection of CagA cytotoxin of Helicobacter pylori].
OBJECTIVE
To obtain and study of immunochemical characteristics of monoclonal antibodies (MAbs) to CagA cytotoxin of Helicobacter pylori employing recombinant fragments of CagA protein.


METHODS
Standard methods of construction and selection of hybridomas, different variants of immunoenzyme analysis and immunoblotting were used. Molecular genotyping of H. pylori cultures by amplification of cagA gene fragments was performed.


RESULTS
Panel of MAbs recognizing 4 different linear epitopes on the CagA molecule, three of which are localized in conservative parts of cytotoxin and one--in variable region of CagA, was developed. On the basis of two obtained antibodies, system of two-center immunoenzyme assay for quantitative detection of CagA protein which is characterized by high sensitivity and specificity, was developed. Obtained MAbs allow to differentiate CagA-positive and CagA-negative strains of H. pylori by immunochemical methods.


CONCLUSIONS
Employing pure recombinant fragments of CagA protein, first panel of MAbs to CagA cytotoxin of H. pylori was developed and characterized. Obtained MAbs open perspectives for study of the H. pylori cytotoxin molecule and construction of immunodiagnostic assays aimed on detection of CagA antigen.
Klimovich
A V
AV
Griazeva
I V
IV
Samoĭlovich
M P
MP
Terekhina
L A
LA
Krutetskaia
I Iu
IIu
Klimovich
V B
VB
D000818:Animals; D017849:Animals, Inbred Strains; D000907:Antibodies, Bacterial / Q000276:immunology*; D000911:Antibodies, Monoclonal / Q000276:immunology*; D000918:Antibody Specificity; D000942:Antigens, Bacterial / Q000032:analysis* / Q000276:immunology; D001426:Bacterial Proteins / Q000032:analysis* / Q000276:immunology; D002678:Chimera; D003603:Cytotoxins / Q000032:analysis* / Q000276:immunology; D018604:Epitope Mapping; D005455:Fluorescent Antibody Technique; D016481:Helicobacter Infections / Q000175:diagnosis*; D016480:Helicobacter pylori / Q000276:immunology* / Q000302:isolation & purification; D006801:Humans; D016056:Immunodominant Epitopes / Q000235:genetics / Q000276:immunology; D051379:Mice; D008815:Mice, Inbred Strains; D010446:Peptide Fragments / Q000276:immunology / Q000302:isolation & purification; D011994:Recombinant Proteins / Q000235:genetics / Q000276:immunology; D012680:Sensitivity and Specificity
D000907:Antibodies, Bacterial; D000911:Antibodies, Monoclonal; D000942:Antigens, Bacterial; D001426:Bacterial Proteins; D003603:Cytotoxins; D016056:Immunodominant Epitopes; D010446:Peptide Fragments; D011994:Recombinant Proteins; C080866:cagA protein, Helicobacter pylori