Study On Preparation And Application Of Monoclonal Antibody Against Penicillin

Penicillins have being the most widely used antibiotics for infection therapy as nontoxic and high effect curative drugs. However, the high allergic reaction rate of penicillins limits their wide useage.lt has been well known that the allergens were not penicillins themselves . Extrinsic imputities such as benzylpenicilloyl protein peptide and/or intrinsic imputity such as polymers are the main causative substances. Penicllin or ampenicillin sometimes were added in biological products manufacturing process .which may form benzylpenicilloyl protein contaminated in final products and have potential danger for penicillin allergic patients.The aim of this study is to develop monoclonal antibody against penicillin and to establish immunoassay method to detect benzylpenicilloyl protein.In this study, we prepared complete antigen by conjugating penicillin with carrier protein to immune BALB/c mice. The hybridoma technique was used to prepare McAbs against penicillin, and the hybridoma culture supernatants were detected by indirect ELISA. Nine hybridoma cell strains stably secreting anti-penicillin McAbs were obtained. These antibodies secreted by those hybridoma cell strains were all lgG1 (k) , of which ascites titers were between1:10~3 to 1:10~8. 5 strains had relative high affinities. All anti-penicillin McAbs specifically recognized penicillin without cross-reaction to carrier protein. Competitive inhibition ELISA assay was established to evaluate the crossreactivity of these McAbs , they all have crossreactivity with penicillins...