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Title: Antifouling Lipid Membranes over Protein A for Orientation-Controlled Immunosensing in Undiluted Serum and Plasma.

Authors: McKeating, Kristy S; Hinman, Samuel S; Rais, Nor Akmaliza; Zhou, Zhiguo; Cheng, Quan

Published In ACS Sens, (2019 07 26)

Abstract: An important advance in biosensor research is the extension and application of laboratory-developed methodologies toward clinical diagnostics, though the propensity toward nonspecific binding of materials in clinically relevant matrices, such as human blood serum and plasma, frequently leads to compromised assays. Several surface chemistries have been developed to minimize nonspecific interactions of proteins and other biological components found within blood and serum samples, though these often exhibit substantially variable outcomes. Herein we report a surface chemistry consisting of a charged-matched supported lipid membrane that has been tailored to form over a gold surface functionalized with protein A. Fine tuning of the interfacial charge of this membrane, along with rational selection of a backfilling self-assembled monolayer, allows for high surface coverage with retention of orientation-controlled capture antibody attachment. We demonstrate using surface-plasmon resonance (SPR) that this highly charged lipid membrane is antifouling, allowing for complete removal of nonspecific human serum and plasma components using only a mild buffer rinse, which we attribute to unique steric interactions with the underlying surface. Furthermore, this surface chemistry is successfully applied for specific detection of IgG and cholera toxin in undiluted human biofluids with negligible sacrifice of SPR signal compared to buffered analysis. This novel lipid membrane interface over protein A may open new avenues for direct biosensing of disease markers within clinical samples.

PubMed ID: 31262175 Exiting the NIEHS site

MeSH Terms: Animals; Antibodies, Immobilized/immunology; Cholera Toxin/blood*; Cholera Toxin/immunology; Gold/chemistry; Humans; Immunoassay/methods; Immunoglobulin Fab Fragments/immunology; Immunoglobulin G/blood*; Immunoglobulin G/immunology; Membranes, Artificial*; Mice; Phosphatidylcholines/chemistry; Phosphatidylglycerols/chemistry; Proof of Concept Study; Staphylococcal Protein A/chemistry*

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