Please use this identifier to cite or link to this item: http://hdl.handle.net/10397/36228
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Title: Characterization of the commercially-available fluorescent Chloroquine-BODIPY Conjugate, LynxTag-CQ(GREEN), as a marker for Chloroquine resistance and uptake in a 96-Well Plate Assay
Authors: Loh, CCY
Suwanarusk, R
Lee, YQ
Chan, KWK
Choy, KY 
Renia, L
Russell, B
Lear, MJ
Nosten, FH
Tan, KSW
Chow, LMC 
Issue Date: 2014
Source: PLoS one, 2014, v. 9, no. 10, e110800
Abstract: Chloroquine was a cheap, extremely effective drug against Plasmodium falciparum until resistance arose. One approach to reversing resistance is the inhibition of chloroquine efflux from its site of action, the parasite digestive vacuole. Chloroquine accumulation studies have traditionally relied on radiolabelled chloroquine, which poses several challenges. There is a need for development of a safe and biologically relevant substitute. We report here a commercially-available green fluorescent chloroquine-BODIPY conjugate, LynxTag-CQ(GREEN), as a proxy for chloroquine accumulation. This compound localized to the digestive vacuole of the parasite as observed under confocal microscopy, and inhibited growth of chloroquine-sensitive strain 3D7 more extensively than in the resistant strains 7G8 and K1. Microplate reader measurements indicated suppression of LynxTag-CQ(GREEN) efflux after pretreatment of parasites with known reversal agents. Microsomes carrying either sensitiveor resistant-type PfCRT were assayed for uptake; resistant-type PfCRT exhibited increased accumulation of LynxTag-CQ(GREEN), which was suppressed by pretreatment with known chemosensitizers. Eight laboratory strains and twelve clinical isolates were sequenced for PfCRT and Pgh1 haplotypes previously reported to contribute to drug resistance, and pfmdr1 copy number and chloroquine IC(50)s were determined. These data were compared with LynxTag-CQ(GREEN) uptake/fluorescence by multiple linear regression to identify genetic correlates of uptake. Uptake of the compound correlated with the logIC(50) of chloroquine and, more weakly, a mutation in Pgh1, F1226Y.
Publisher: Public Library of Science
Journal: PLoS one 
EISSN: 1932-6203
DOI: 10.1371/journal.pone.0110800
Rights: © 2014 Loh et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
The following publication: Loh CCY, Suwanarusk R, Lee YQ, Chan KWK, Choy K-Y, Rénia L, et al. (2014) Characterization of the Commercially-Available Fluorescent Chloroquine-BODIPY Conjugate, LynxTag-CQGREEN, as a Marker for Chloroquine Resistance and Uptake in a 96-Well Plate Assay. PLoS ONE 9(10): e110800 is available at https://doi.org/10.1371/journal.pone.0110800
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