Development of Monoclonal Antibodies against Helicobacter pylori and Their Applications to Immunological Assays
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Abstract:
Helicobacter pylori (Hp) is a gram-negative, microaerophilic bacterium found in the human stomach and duodenum, and long-term H. pylori infection is linked to the development of gastritis, gastric and duodenal ulcers, and stomach cancer. H. pylori infection is mainly acquired during childhood, and its spontaneous clearance is rare. Its prevalence among adults can be effectively reduced by early diagnosis and antibiotic therapy. In this study, a standard H. pylori strain (ATCC 43504) was cultured in vitro, the lysate was used as an antigen to immunize BALB/c mice, and a hybridoma cell line was prepared using conventional techniques. In total, 21 hybridoma cell lines that stably secreted anti-H. pylori monoclonal antibodies (mAbs) were obtained based on indirect enzyme-linked immunosorbent assay (ELISA) screening and multiple rounds of limiting dilution subcloning. Ascites were prepared and mAbs were purified using ammonium sulfate precipitation methods. The pairing of mAbs and sample detection were conducted by colloidal gold immunochromatographic assays (GICAs), and multiple mAb pairs with high sensitivity and specificity were obtained. Among them, the limits of detection (LODs) of the lysates from two antibody pairs consisting of capture (mAb 2-18E1) and detection (mAb 2-9H5 and mAb 2-17E9) antibodies were 25 ng/mL. These results indicated that the GICA method can provide a basis for the development of reagents to detect H. pylori antigens.
