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Metal Affinity-Enabled Capture and Release Antibody Reagents Generate a Multiplex Biomarker Enrichment System that Improves Detection Limits of Rapid Diagnostic Tests
journal contribution
posted on 2017-08-30, 00:00 authored by Westley S. Bauer, Christopher P. Gulka, Lidalee Silva-Baucage, Nicholas M. Adams, Frederick R. Haselton, David W. WrightMulti-antigen rapid diagnostic tests
(RDTs) are highly informative,
simple, mobile, and inexpensive, making them valuable point-of-care
(POC) diagnostic tools. However, these RDTs suffer from several technical
limitationsthe most significant being the failure to detect
low levels of infection. To overcome this, we have developed a magnetic
bead-based multiplex biomarker enrichment strategy that combines metal
affinity and immunospecific capture to purify and enrich multiple
target biomarkers. Modifying antibodies to contain histidine-rich
peptides enables reversible loading onto immobilized metal affinity
magnetic beads, generating a novel class of antibodies coined “Capture
and Release” (CaR) antibody reagents. This approach extends
the specificity of immunocapture to metal affinity magnetic beads
while also maintaining a common trigger for releasing multiple biomarkers.
Multiplex biomarker enrichment is accomplished by adding magnetic
beads equipped with CaR antibody reagents to a large sample volume
to capture biomarkers of interest. Once captured, these biomarkers
are magnetically purified, concentrated, and released into a RDT-compatible
volume. This system was tailored to enhance a popular dual-antigen
lateral flow malaria RDT that targets Plasmodium falciparum histidine-rich protein-II (HRPII) and Plasmodium lactate dehydrogenase (pLDH). A suite of pLDH CaR antibody reagents were synthesized, characterized,
and the optimal CaR antibody reagent was loaded onto magnetic beads
to make a multiplex magnetic capture bead that simultaneously enriches pLDH and HRPII from Plasmodium falciparum parasitized blood samples. This system achieves a 17.5-fold improvement
in the dual positive HRPII/pan-pLDH detection limits
enabling visual detection of both antigens at levels correlating to
5 p/μL. This front-end sample processing system serves as an
efficient strategy to improve the sensitivity of RDTs without the
need for modifications or remanufacturing.
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p LDH CaR antibody reagentsbeadsPlasmodium lactate dehydrogenaseMultiplex Biomarker Enrichment SystemMultiplex biomarker enrichmentCaR antibody reagentRapid Diagnostic Tests Multi-antigenflow malaria RDTRelease Antibody Reagents GenerateHRPIImetal affinityp LDHCaR antibody reagentsbead-based multiplex biomarker enrichment strategyPlasmodium falciparum parasitized blood samplesPOCsample processing systemtargets Plasmodium falciparum histidine-rich protein-IImetal Affinity-Enabled Capture
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