Preparation And Application Of The Monoclonal Antibody Against Baicalin

Monoclonal antibody(MAb) technology has been widely applied in bio-medicinal practice since it was established.Theoretically,the corresponding monoclonal antibodies against its bio-active components of traditional Chinese medcines(TCM) can produced by hybridoma technique.Therefore,these MAb can be used to determine and /or analyse the bio-active components from TCM row materials,to obtain high-purity bio-actives through affinity chromatography,to monitor or control the quality of TCM products in manufacturing process,doubtlessly, can also be used in such as phytochemistry,pharmacology and pharceurical researches,etc.Baicalin(BCL) is one of major chemical components of Scutellaria baicalensis Georgi,a very-common-used TCM herbal source.Recent researches on BCL showed that the baicalin has lots of bio-active characteristics,such as,strong anti-oxidation ability,reducing the injuries of heart and cerebral ischemia,improving in diabetic deseas complication,protecting liver,regulating blood-fat,dilating vessels,preventing athero-sclerosis and depressurizing.The most promissing pharmacologcal effects including anti-tumor activity,anti-HIV and influenza virus,as a natural inhibitor of reverse transcriptase.This work aimed at:1.Preparation of the anti-BCL MAbs;The two BCL-MAbs obtained,B11F and F8C,have the characteristics as follows:sensitivity are 30ng/ml and 20ng/ml respectively;affinity constants are 2.975×10^-10 mol/L and 3.265×10^-10 mol/L respectively;the specificity of the two monoclonal antibodies is very high and they basically do not have cross-reactions with Ursolic acid,Daidzein,Rutin and Glycyrrhizic acid;the cross-reaction rates with Baicalein are 7.8%and 5.5%.2.Establishment of an enzyme-linked immunosorbent assay(ELISA) in order to to detemine the BCL concentration of raw Scutellaria materia medica,and Chinese patent medicines with ingredient of BCL.The results we obtained showed that the established ELISA assay using anti-BCL MAb is much more fast,sensitive and accurate,comparing with concentional HPLC determination.It also suggests higher feasiblity in other relavant bio-medical researches including medicinal chemistry,pharmaceutical science and pharmacology.