title: Development of a sensitive monoclonal antibody-based enzyme-linked immunosorbent assay for the antimalaria active ingredient artemisinin in the Chinese herb Artemisia annua L.
abstract: Artemisinin is an endoperoxide sesquiterpene lactone isolated from the Chinese medicinal plant Artemisia annua L. It has been widely used in South-East Asia and Africa as an effective drug against sensitive and multidrug-resistant Plasmodium falciparum malaria. A monoclonal antibody (mAb), designated as 3H2, was generated with artesunate-bovine serum albumin conjugate as the immunogen. mAb 3H2 was used to develop a highly sensitive and specific indirect competitive enzyme-linked immunosorbent assay (icELISA) for artemisinin. The concentration of analyte producing 50% of inhibition (IC(50)) and the working range of the icELISA were 1.3 and 0.2-5.8 ng/mL, respectively. The mAb 3H2 recognized the artemisinin analogs artesunate, dihydroartemisinin, and artemether with cross-reactivity of 650%, 57%, and 3%, respectively, but negligibly recognized deoxyartemisinin and the artemisinin precursors arteannuin B and artemisinic acid. The average recoveries of artemisinin fortified in A. annua samples at concentrations from 156 to 5,000 microg/g determined by icELISA ranged from 91% to 98%. The icELISA was applied for the determination of artemisinin in different wild A. annua samples and the results were confirmed by high-performance liquid chromatography (HPLC) analysis. The correlation coefficient (R(2)) between the two assays was larger than 0.99, demonstrating a good agreement between the icELISA and HPLC results. This ELISA is suitable for quality assurance of A. annua L. materials.
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University, Beijing, 100193, China.
authors.forename: Su-Ping
authors.initials: SP
authors.lastname: He
authors.forename: Gui-Yu
authors.initials: GY
authors.lastname: Tan
authors.forename: Gang
authors.initials: G
authors.lastname: Li
authors.forename: Wei-Ming
authors.initials: WM
authors.lastname: Tan
authors.forename: Tie-Gui
authors.initials: TG
authors.lastname: Nan
authors.forename: Bao-Min
authors.initials: BM
authors.lastname: Wang
authors.forename: Zhao-Hu
authors.initials: ZH
authors.lastname: Li
authors.forename: Qing X
authors.initials: QX
authors.lastname: Li
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology*; D019306:Artemisia / Q000737:chemistry*; D037621:Artemisinins / Q000032:analysis*; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods* / Q000592:standards; D007106:Immune Sera; D012680:Sensitivity and Specificity
chemical_list: D000911:Antibodies, Monoclonal; D037621:Artemisinins; D007106:Immune Sera; C031327:artemisinin