title: Development of a Specific Monoclonal Antibody for the Quantification of Artemisinin in Artemisia annua and Rat Serum.
abstract: Artemisinin, extracted from Artemisia annua, and its derivatives are important frontline antimalarials. To produce specific antibodies for the detection and quantification of artemisinin, artemisinin was transformed to 9-hydroxyartemisinin by microbial fermentation, which was used to prepare a 9-succinate artemisinin hapten for conjugation with ovalbumin. A monoclonal antibody (mAb), designated as 3H7A10, was selected from hybridoma cell lines which showed high specificity to artemisinin. No competitive inhibition was observed with artesunate, dihydroartemisinin, and artemether for up to 20,000 ng mL(-1). An indirect competitive enzyme-linked immunosorbent assay (icELISA) was developed, which showed a concentration causing 50% of inhibition (IC50) for artemisinin as 2.6 ng mL(-1) and a working range of 0.6-11.5 ng mL(-1). The icELISA was applied for the quantification of artemisinin in crude extracts of wild A. annua and the study of pharmacokinetics of artemisinin in rat serum after intraperitoneal injection. The results were highly correlated with those determined by HPLC-UV analysis (R(2) = 0.9919). In comparison with reported antiartemisinin mAbs which have broad cross-reactivity with other artemisinin derivatives, the high specificity of 3H7A10 for artemisinin will enable development of methods for quantification of artemisinin in Artemisia plants and antimalarial drugs such as Arco and for pharmacokinetic studies.
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University , 100193 Beijing, China.
authors.forename: Suqin
authors.initials: S
authors.lastname: Guo
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University , 100193 Beijing, China.
authors.forename: Yongliang
authors.initials: Y
authors.lastname: Cui
authors.affiliation: College of Agronomy and Biotechnology, Yunnan Agricultural University , Kunming, Yunnan 650201, China.
authors.forename: Kunbi
authors.initials: K
authors.lastname: Wang
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University , 100193 Beijing, China.
authors.forename: Wei
authors.initials: W
authors.lastname: Zhang
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University , 100193 Beijing, China.
authors.forename: Guiyu
authors.initials: G
authors.lastname: Tan
authors.affiliation: College of Agronomy and Biotechnology, China Agricultural University , 100193 Beijing, China.
authors.forename: Baomin
authors.initials: B
authors.lastname: Wang
authors.affiliation: Department of Entomology, Pennsylvania State University , University Park, Pennsylvania 16802, United States.
authors.forename: Liwang
authors.initials: L
authors.lastname: Cui
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology*; D031172:Artemisia annua / Q000737:chemistry; D037621:Artemisinins / Q000032:analysis* / Q000097:blood / Q000276:immunology; D004797:Enzyme-Linked Immunosorbent Assay; D006825:Hybridomas; D008807:Mice, Inbred BALB C; D017207:Rats, Sprague-Dawley
chemical_list: D000911:Antibodies, Monoclonal; D037621:Artemisinins; C031327:artemisinin
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