Preparation of biotinylated anti-cytokine monoclonal antibodies
Abstract：Objective To explore a method of preparation for biotinylatedmonoclonal antibodies． Methods Rat anti-mouse IL-5， IL-4 and IFN-γ were purified by ultra filtration， precipitated with saturated ammonium sulfate further purified by chromatography with the protein G-sepharose affinity column from the hybridoma cell culture supernatants． We added 80 μg of the biotinylation reagent per 1 mg of the monoclonal antibody， and incubated them at room temperature for 60 min． After that， we applied the monoclonal antibody sample to the Sephadex G-25 column and got the purified biotinylated monoclonal antibody． The antibodies were tested the activities through the Avidin-biotin ELISA． Results The results showed that the antibodies kept high biological activities and sensitivities in detecting the sera from IL-5 transgenic mice and non-transgenic mice by ELISA． Conclusion The method is simple and stable．
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Publication Date：2001-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：3( 222-224 )