Preparation of a Novel Monoclonal Antibody against the Antimalarial Drugs, Artemisinin and Artesunate

Tanaka, Hiroyuki; Putalun, Waraporn; De-Eknamkul, Wanchai; Matangkasombut, Oraphan; Shoyama, Yukihiro

doi:10.1055/s-2007-981560

Cited by 19 publications

(18 citation statements)

References 24 publications

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“…To obtain ART-BSA conjugate as the immunogen of ART, Tanaka et al simultaneously incubated ART, BSA, and EDC [15]. However, this method had the disadvantage that self-coupling of the BSA molecules might occur.…”

Section: Discussionmentioning

confidence: 99%

“…By contrast, enzyme-linked immunosorbent assays (ELISAs) are generally very simple and specific methods for quantitative measurements of drugs. Ferreira and Janick and Tanaka et al produced polyclonal antibodies (pAbs) and monoclonal antibodies (mAbs) to ART [14, 15], respectively, and developed an indirect competitive ELISA for detection of ARM in botanical samples. Subsequently, Wang et al developed an indirect competitive ELISA of ART using anti-ART mAb produced by He et al and applied the assay for the quantification of ART tablets [16, 17].…”

Section: Introductionmentioning

confidence: 99%

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Development of a simple and specific direct competitive ELISA for the determination of artesunate using an anti-artesunate polyclonal antiserum

Mitsui

2016

Trop Med Health

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BackgroundSince artesunate (ART) became a vital component of artemisinin (ARM)-based combination therapies for the treatment for malaria, counterfeit ART drugs have spread in regions of Southeast Asia and Africa. The consumption of counterfeit ART drugs has resulted in the death of many patients. Thus, evaluating the quality of ART drugs is needed. There are several methods for quantitating the ART content in tablets, the most common being a high-performance liquid chromatography. However, that method is hampered by the need for expensive equipment and a rather time-consuming process of extraction. By contrast, enzyme-linked immunosorbent assays (ELISAs) are faster and much less expensive, and they require less sample preparation than the above method. The objective of the present study was to establish a simple and specific direct competitive ELISA for the determination of ART concentrations using an anti-ART polyclonal antibody (pAb).ResultsAnti-ART pAb was raised in mice, and ART-horseradish peroxidase (HRP) conjugate was produced. A direct competitive ELISA was performed by simultaneously incubating ART and the ART-HRP conjugate with the anti-ART pAb over a second antibody. Subsequently, the enzyme activity of the remaining ART-HRP conjugate was measured. The intra- and inter-assay coefficients of variation of the ELISA were less than 10 % in the range of 0.3 to 30 ng/ml with a detection limit of 0.1 ng/ml. The cross-reactivities of the anti-ART pAb with ARM and dihydroartemisinin were 0.12 and 0.04 %, respectively, and those with other antimalarial drugs were negligible. Furthermore, the recovery of 10 or 50 ng/ml ART added to the drug tablet solutions containing an expected amount of 10 ng/ml was estimated by the ELISA. The recovery of the ART amount ranged between 98 and 106 %, with coefficient variations of less than 7.0 %.ConclusionsThe present ELISA is a simple and specific method for the determination of ART concentrations. Thus, this ELISA can be used to identify ART counterfeits and substandard drugs and to quantify the ART drugs.

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Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Development of a simple and specific direct competitive ELISA for the determination of artesunate using an anti-artesunate polyclonal antiserum

Mitsui

2016

Trop Med Health

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“…We have developed quality control methods for preparing AM from an Artemisia spp. selected for its ability to produce high yields of AM [34].…”

Section: Am Asmentioning

confidence: 99%

“…An icELISA using anti-AM mAb 1C1 was developed based on the general format that we developed previously [34]. The free mAb 1C1 after competition with A M is bound to polystyrene microtiter plates precoated with AM-HSA (1 g/mL) in the ELISA.…”

Section: Evaluation and Application Of Icelisa Using Anti-am Mabmentioning

confidence: 99%

Immunochemical Analysis of the Antimalarial Drugs Artemisinin and Artesunate

et al. 2012

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Abstract:We prepared a monoclonal antibody (mAb 1C1) showing specificity for artemisinin (AM) and artesunate (AS), and we developed an indirect competitive enzymelinked immunosorbent assay (icELISA) using this novel mAb. Moreover, we prepared a recombinant antibody derived from mAb 1C1 in order to overcome insufficient mAb production by hybridoma culture. A recombinant antigen-binding fragment (Fab) was easily constructed using antibody manipulation technologies and was produced by microorganisms in high yield. We herein review immunochemical approaches for analysis of the antimalarial drugs AM and AS that were able to yield analysis results for multiple samples in a short period of time using simple and reliable protocols.

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“…In the course of a pluridisplinary program aiming to design a versatile mobile laboratory for onsite analysis, we were interested in the development of an enzyme immunoassay for DHA in biological fluids. Currently, a few studies report the preparation of monoclonal or polyclonal antibodies against artemisinin mainly for its quantification in crude plant extracts of Artemisia annua, a Chinese herbal medicine from which it is extracted [10][11][12] or in anti-malarial drugs for quality controls [13]. However, no immunoassay for DHA plasma analysis with a sensitivity, rapidity, and cost-effectiveness at least equal to those of LC methods has been published to date.…”

Section: Introductionmentioning

confidence: 99%

Development of sensitive direct chemiluminescent enzyme immunoassay for the determination of dihydroartemisinin in plasma

et al. 2015

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Despite significant progress in prevention and therapy, malaria is still one of the world's leading major diseases due to its high morbidity and mortality. Recommended treatments by the World Health Organization include the use of artemisinin and artemisinin derivative-based combination therapies. To allow efficient patient monitoring during antimalarial therapy without the use of expensive apparatus, we developed a sensitive direct chemiluminescent enzyme immunoassay for the determination of dihydroartemisinin in biological fluids. To produce specific antibodies against dihydroartemisinin (DHA), a synthetic DHA derivative was coupled to bovine serum albumin as the immunogen. In parallel, a new, rapid, and efficient procedure to covalently link glycoprotein to all amine-containing molecules has been established and the enzyme tracer was prepared by chemically coupling the DHA derivative in combination with SBP rather than the more commonly used HRP. It allowed us to develop, after optimization of the luminescent reagent, a sensitive and stable luminescent EIA, with a LLOQ of 90 pg mL(-1). This assay compares favorably with the most efficient HPLC methods previously reported with a LLOQ close to 1 ng mL(-1) and shows good precision and efficiency since recovery from human plasma spiked with DHA ranged between 91 and 103%, with coefficients of variation of <13%. To date, no immunoassay for DHA has been applied to plasma analysis and this EIA should be very useful in all clinical laboratories for rapid and cost-effective analysis.

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Preparation of a Novel Monoclonal Antibody against the Antimalarial Drugs, Artemisinin and Artesunate

Cited by 19 publications

References 24 publications

Development of a simple and specific direct competitive ELISA for the determination of artesunate using an anti-artesunate polyclonal antiserum

Development of a simple and specific direct competitive ELISA for the determination of artesunate using an anti-artesunate polyclonal antiserum

Immunochemical Analysis of the Antimalarial Drugs Artemisinin and Artesunate

Development of sensitive direct chemiluminescent enzyme immunoassay for the determination of dihydroartemisinin in plasma

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