Development of a specific monoclonal antibody-based ELISA to measure the artemether content of antimalarial drugs.
Artemether is one of the artemisinin derivatives that are active ingredients in antimalarial drugs. Counterfeit and substandard antimalarial drugs have become a serious problem, which demands reliable analytical tools and implementation of strict regulation of drug quality. Structural similarity among artemisinin analogs is a challenge to develop immunoassays that are specific to artemisinin derivatives. To produce specific antibodies to artemether, we used microbial fermentation of artemether to obtain 9-hydroxyartemether, which was subsequently used to prepare a 9-O-succinylartemether hapten for conjugation with ovalbumin as the immunogen. A monoclonal antibody (mAb), designated as 2G12E1, was produced with high specificity to artemether. 2G12E1 showed low cross reactivities to dihydroartemisinin, artemisinin, artesunate and other major antimalarial drugs. An indirect competitive enzyme linked immunosorbent assay (icELISA) developed showed a concentration causing 50% of inhibition for artemether as 3.7 ng mL⁻¹ and a working range of 0.7-19 ng mL⁻¹. The icELISA was applied for determination of artemether content in different commercial drugs and the results were comparable to those determined by high-performance liquid chromatography analysis. In comparison with reported broad cross activity of anti-artemisinin mAbs, the most notable advantage of the 2G12E1-based ELISA is its high specificity to artemether only.
College of Agronomy and Biotechnology, China Agricultural University, Beijing, China.
Suqin
S
Guo
Yongliang
Y
Cui
Lishan
L
He
Liang
L
Zhang
Zhen
Z
Cao
Wei
W
Zhang
Rui
R
Zhang
Guiyu
G
Tan
Baomin
B
Wang
Liwang
L
Cui
D000818:Animals; D058846:Antibodies, Monoclonal, Murine-Derived / Q000096:biosynthesis / Q000737:chemistry*; D000918:Antibody Specificity; D000962:Antimalarials / Q000032:analysis* / Q000302:isolation & purification; D000077549:Artemether; D037621:Artemisinins / Q000032:analysis* / Q000302:isolation & purification; D001667:Binding, Competitive; D004339:Drug Compounding; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006825:Hybridomas; D051379:Mice; D008807:Mice, Inbred BALB C; D011485:Protein Binding
D058846:Antibodies, Monoclonal, Murine-Derived; D000962:Antimalarials; D037621:Artemisinins; D000077549:Artemether