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Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations
BACKGROUND: In Central and South America and Eastern and Southern Africa, Plasmodium vivax infections accounts for 71-81% and 5% of malaria cases, respectively. In these areas, chloroquine (CQ) remains the treatment of choice for P. vivax malaria. In addition, CQ has recently proven to be an effecti...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3170649/ https://www.ncbi.nlm.nih.gov/pubmed/21864375 http://dx.doi.org/10.1186/1475-2875-10-249 |
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author | Khalil, Insaf F Alifrangis, Michael Recke, Camilla Hoegberg, Lotte C Ronn, Anita Bygbjerg, Ib C Koch, Claus |
author_facet | Khalil, Insaf F Alifrangis, Michael Recke, Camilla Hoegberg, Lotte C Ronn, Anita Bygbjerg, Ib C Koch, Claus |
author_sort | Khalil, Insaf F |
collection | PubMed |
description | BACKGROUND: In Central and South America and Eastern and Southern Africa, Plasmodium vivax infections accounts for 71-81% and 5% of malaria cases, respectively. In these areas, chloroquine (CQ) remains the treatment of choice for P. vivax malaria. In addition, CQ has recently proven to be an effective HIV-1 therapeutic agent. There is a dire need to continue monitoring quality of CQ as there is a major influx of substandard and fake formulations into malaria-endemic countries. The use of fake/substandard drugs will result in sub-therapeutic levels endangering the patient and possibly select for parasite resistance. The aim of this study was to develop an inexpensive, simple antibody-based ELISA to measure CQ concentrations in tablets and in plasma. METHODS: A monoclonal antibody (MAb) that reacts with the N-side chain of the CQ molecule was prepared by use of a CQ analogue. A specific and reliable ELISA for detection of CQ was developed. The developed assay was validated by measuring CQ in tablets sold in Denmark, India and Sudan. Furthermore, kinetics of CQ concentrations in plasma of four volunteers, who ingested two tablets of Malarex(® )containing, 250 mg CQ base, were measured before drug intake, three hours later and thereafter at days 1, 3, 7, 14, 21 and 28. The same plasma samples were simultaneously measured by high performance liquid chromatography (HPLC). RESULTS: The ELISA proved an easy-to-handle and very sensitive tool for the detection of CQ with a lower limit of detection at 3.9 ng/ml. ELISA levels of CQ in plasma showed high agreement with the levels obtained by HPLC (r = 0.98). The specificity in the negative control group was 100%. CONCLUSION: The developed ELISA can be used for quality screening of CQ in pharmaceutical formulations and for drug monitoring in malaria and in other infectious diseases, such as HIV, where CQ proved to be an effective therapeutic agent. The methodology has been exploited to develop monoclonal antibodies for the drugs used in artemisinin-based combination therapy (ACT). |
format | Online Article Text |
id | pubmed-3170649 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-31706492011-09-11 Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations Khalil, Insaf F Alifrangis, Michael Recke, Camilla Hoegberg, Lotte C Ronn, Anita Bygbjerg, Ib C Koch, Claus Malar J Methodology BACKGROUND: In Central and South America and Eastern and Southern Africa, Plasmodium vivax infections accounts for 71-81% and 5% of malaria cases, respectively. In these areas, chloroquine (CQ) remains the treatment of choice for P. vivax malaria. In addition, CQ has recently proven to be an effective HIV-1 therapeutic agent. There is a dire need to continue monitoring quality of CQ as there is a major influx of substandard and fake formulations into malaria-endemic countries. The use of fake/substandard drugs will result in sub-therapeutic levels endangering the patient and possibly select for parasite resistance. The aim of this study was to develop an inexpensive, simple antibody-based ELISA to measure CQ concentrations in tablets and in plasma. METHODS: A monoclonal antibody (MAb) that reacts with the N-side chain of the CQ molecule was prepared by use of a CQ analogue. A specific and reliable ELISA for detection of CQ was developed. The developed assay was validated by measuring CQ in tablets sold in Denmark, India and Sudan. Furthermore, kinetics of CQ concentrations in plasma of four volunteers, who ingested two tablets of Malarex(® )containing, 250 mg CQ base, were measured before drug intake, three hours later and thereafter at days 1, 3, 7, 14, 21 and 28. The same plasma samples were simultaneously measured by high performance liquid chromatography (HPLC). RESULTS: The ELISA proved an easy-to-handle and very sensitive tool for the detection of CQ with a lower limit of detection at 3.9 ng/ml. ELISA levels of CQ in plasma showed high agreement with the levels obtained by HPLC (r = 0.98). The specificity in the negative control group was 100%. CONCLUSION: The developed ELISA can be used for quality screening of CQ in pharmaceutical formulations and for drug monitoring in malaria and in other infectious diseases, such as HIV, where CQ proved to be an effective therapeutic agent. The methodology has been exploited to develop monoclonal antibodies for the drugs used in artemisinin-based combination therapy (ACT). BioMed Central 2011-08-24 /pmc/articles/PMC3170649/ /pubmed/21864375 http://dx.doi.org/10.1186/1475-2875-10-249 Text en Copyright ©2011 Khalil et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. |
spellingShingle | Methodology Khalil, Insaf F Alifrangis, Michael Recke, Camilla Hoegberg, Lotte C Ronn, Anita Bygbjerg, Ib C Koch, Claus Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations |
title | Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations |
title_full | Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations |
title_fullStr | Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations |
title_full_unstemmed | Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations |
title_short | Development of ELISA-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations |
title_sort | development of elisa-based methods to measure the anti-malarial drug chloroquine in plasma and in pharmaceutical formulations |
topic | Methodology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3170649/ https://www.ncbi.nlm.nih.gov/pubmed/21864375 http://dx.doi.org/10.1186/1475-2875-10-249 |
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