Establishment Of Monoclonal Antibody-based Immunoassay Of Two Bio-Active Components From Some Compound Prescriptions Of Traditional Chinese Medicines

In this experiment,two monoclonal antibodies(MAbs) against major bioactive components of glycyrrhizic acid(GA) and paoniflorin(PF) from the compound prescription of traditional Chinese medicine "Shaoyao Gancao decoction" were prepared based on the hybridoma technique and purified according to protein G chromatography.A sensitive enzyme-linked.......(ELISA) using these two anti-GA and anti-PF MAbs to quantify the glycyrrhizin and paoniflorin was established.The ELISA method was also successively used to determine the contents of GA and PF in the extracts of ten market-available traditional Chinese drugs.The main steps cover the following:1.Preparation of artificial antigens of GA-HSA and PF-HSA;2.Immunization of animals;3.Cloning and selecting of antibody secreted cells by hybridoma technology;4.Scaling-up the MAb-producing cells;5.Harvesting and purification of the target MAbs;6.Characterizing of the prepared MAbs;7.Establishment of ELISA based the two MAbs;8.Determination of GA and PF from other ten Chinese drugs extracts;9.Comparison the ELISA results with HPLC's.The results of the ELISA and HPLC detection were in high agreement(≤5%),suggesting that the ELISA method based on MAbs to detect the bioactive components of Chinese traditional medicines was feasible and effective.This method also provided an alternative and comparative way to detect the other bioactive components of compound prescription of traditional Chinese medicines.