Development of Escherichia coli Asparaginase II for Immunosensing: A Trade-Off between Receptor Density and Sensing Efficiency

ACS Omega
David M CharbonneauJoelle N Pelletier

Abstract

The clinical success of Escherichia colil-asparaginase II (EcAII) as a front line chemotherapeutic agent for acute lymphoblastic leukemia (ALL) is often compromised because of its silent inactivation by neutralizing antibodies. Timely detection of silent immune response can rely on immobilizing EcAII, to capture and detect anti-EcAII antibodies. Having recently reported the use of a portable surface plasmon resonance (SPR) sensing device to detect anti-EcAII antibodies in undiluted serum from children undergoing therapy for ALL (Aubé et al., ACS Sensors2016, 1 (11), 1358-1365), here we investigate the impact of the quaternary structure and the mode of immobilization of EcAII onto low-fouling SPR sensor chips on the sensitivity and reproducibility of immunosensing. We show that the native tetrameric structure of EcAII, while being essential for activity, is not required for antibody recognition because monomeric EcAII is equally antigenic. By modulating the mode of immobilization, we observed that low-density surface coverage obtained upon covalent immobilization allowed each tetrameric EcAII to bind up to two antibody molecules, whereas high-density surface coverage arising from metal chelation by N- or C-terminal histidine-tag...Continue Reading

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Citations

May 28, 2020·Frontiers in Microbiology·Anupam SinghAmulya K Panda
May 19, 2020·Biomedit︠s︡inskai︠a︡ khimii︠a︡·M V DuminaN N Sokolov

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Methods Mentioned

BETA
surface plasmon resonance
chips
enzyme-linked immunosorbent assay
electrophoresis
Size
ELISA
size exclusion chromatography
circular dichroism
chip
PCR

Software Mentioned

GraphPad Prism
Clone Manager
MATLAB

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