%0 Journal Article
%A Bauer, Westley S.
%A Gulka, Christopher P.
%A Silva-Baucage, Lidalee
%A Adams, Nicholas M.
%A Haselton, Frederick R.
%A Wright, David W.
%D 2017
%T Metal Affinity-Enabled Capture and Release Antibody
Reagents Generate a Multiplex Biomarker Enrichment System that Improves
Detection Limits of Rapid Diagnostic Tests
%U https://acs.figshare.com/articles/journal_contribution/Metal_Affinity-Enabled_Capture_and_Release_Antibody_Reagents_Generate_a_Multiplex_Biomarker_Enrichment_System_that_Improves_Detection_Limits_of_Rapid_Diagnostic_Tests/5425621
%R 10.1021/acs.analchem.7b01513.s001
%2 https://acs.figshare.com/ndownloader/files/9360886
%K p LDH CaR antibody reagents
%K beads
%K Plasmodium lactate dehydrogenase
%K Multiplex Biomarker Enrichment System
%K Multiplex biomarker enrichment
%K CaR antibody reagent
%K Rapid Diagnostic Tests Multi-antigen
%K flow malaria RDT
%K Release Antibody Reagents Generate
%K HRPII
%K metal affinity
%K p LDH
%K CaR antibody reagents
%K bead-based multiplex biomarker enrichment strategy
%K Plasmodium falciparum parasitized blood samples
%K POC
%K sample processing system
%K targets Plasmodium falciparum histidine-rich protein-II
%K metal Affinity-Enabled Capture
%X Multi-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.
%I ACS Publications