Development of high throughput immunoassays for analysing artemisinin & derivatives
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Thematic areas: Health, Lead institution: University of Copenhagen (UCPH)/Rigshospitalet, Centre for Medical Parasitology (CMP), University of Copenhagen (UCPH)/Rigshospitalet, Denmark, Total grant: 1,908,432 DKK Contact person: Insaf Fadi Khalil
During the project period specific monoclonal antibodies (MAbs) have been raised against artesunate (ARS), dihydroartemisinin (DHA), chloroquine (CQ) and amodiaquine (AQ). By using these antibodies, highly sensitive and specific enzyme-linked immunosorbent assays (ELISA) were developed for quantifying the four drugs in pharmaceutical formulations and in plasma. The ELISA system shows a full measuring range from 1 to 750ng/ml in the case of ARS and DHA and from 1 to 500ng/ml in the case of CQ and AQ. The developed ELISAs proved an easy-to-handle and very sensitive tool with lower limit of detections of less than 3ng/ml for all tested drugs. The specificity in the negative control group was 100%. During the project period a reversed phase high performance liquid chromatography method for measuring lumefantrine and its main metabolite in plasma has been developed and evaluated. The method has been transferred to The National Institute of Medical research (Tanga/Tanzania) and it will be used in conjugation with the ongoing pharmacovigilance studies of artemisinins combination therapy. The monoclonal antibodies generated for AQ and CQ are commercially available through the Danish biotechnology company BioPort A/S. For the moment we are seeking fund to transfer the ELISA methods into commercially available dipsticks that could be used as rapid tests for quality control of the tested pharmaceuticals.