Preparing and immunological characterization of monoclonal antibodies against parvalbumin—a major allergen from bighead carps

Objective To prepare,purify and identify monoclonal antibodies against parvalbumin from bighead carps.Methods Using hybridoma and limit dilution technique,the fusion cell strain secreting antibodies stably was screened.The cells were used to induce ascites in mice and monoclonal antibodies were purified using affinity chromatography on immobilized protein A.The subclass of Ig,speciality,titers,and cross-reactvity of the antibdies were detected with indirect enzymelinked immunosorbent assay(ELISA) and western blot.Results Seven cell lines secreting antibodies against bighead carp parvalbumin were prepared.The titers of 5 of 7 monoclonal antibodies(1G1,2B9,1H4,1B7,and 1G3) were higher than 1:400 000 and those of 1B5 and 1A5 were higher than 1:200 000 (P/N > 2.1).Results of western blot indicated that all antibodies could bind to parvalbumin specifically.ELISA showed that the subclass of the Ig was IgG1.1G1,1A5,2B9,1H4,and 1B7 showed specifec binding to parvalbumin,whereas,1G3 and 1A5 had cross-reactivity with allergen from soybean and shrimp (P/N > 2.1).Conclusion This research successfully prepared 5 antibodies with high titers for a detection system of parvalbumin and found the allergen composition reacting to the 2 of the 5 monoclonal antibodies from shrimp and soybean.