Characterization of specific monoclonal antibodies for detection of mefloquine in body fluids.
Specific monoclonal antibodies (MAbs) to mefloquine conjugated to bovine serum albumin (mefloquine-BSA) were produced by hybridoma technology. The mefloquine-BSA was synthesized by converting mefloquine into hemisuccinate followed by convalently linked to bovine serum albumin (BSA) and coupling with N,N' disuccinimidyl carbonate (DSC). The conjugate was purified by Sephadex G-75 gel filtration using 0.01 M PBS pH 7.2. An average of 19.34 molecules of mefloquine were conjugated to each molecule of protein determined by differential UV absorption spectra of hapten and protein carrier. Sixteen monoclones producing antibody specific to mefloquine were screened by indirect ELISA using homologous antigens. The specificity of MAbs was determined by reacting with BSA and the structurally related antimalarial drug, quinine. Three, three, five and two MAbs belonged to IgG1, IgG2a, IgG2b and IgG3, respectively. Most of the MAbs slightly reacted with quinine-BSA due to the closely related structure of mefloquine to quinine. The selected MAb designated 11F9(G5)G9 which showed no cross reaction with quinine-BSA gave high reactivity with blood samples from malaria patients previously treated with mefloquine when compared to normal blood by indirect ELISA. The preliminary results indicated that such specific MAb could be used as antibody probe for detection of mefloquine in biological fluids.
Trisirivanich
S
S
Department of Microbiology and Immunology, Faculty of Tropical Medicine, Mahidol University, Bangkok, Thailand.
Laothavorn
J
J
Na-Bangchang
K
K
Khusmith
S
S
D000911:Antibodies, Monoclonal / Q000097:blood* / Q000276:immunology; D000962:Antimalarials / Q000097:blood* / Q000276:immunology / Q000652:urine*; D002851:Chromatography, High Pressure Liquid; D003429:Cross Reactions; D004797:Enzyme-Linked Immunosorbent Assay; D006801:Humans; D016778:Malaria, Falciparum / Q000097:blood* / Q000188:drug therapy; D015767:Mefloquine / Q000097:blood* / Q000276:immunology / Q000652:urine*; D012680:Sensitivity and Specificity; D012710:Serum Albumin, Bovine / Q000276:immunology; D013785:Thailand
D000911:Antibodies, Monoclonal; D000962:Antimalarials; D012710:Serum Albumin, Bovine; D015767:Mefloquine