Scanning force microscopy and fluorescence microscopy of microcontact printed antibodies and antibody fragments

Langmuir : the ACS Journal of Surfaces and Colloids
John R LaGraff, Quynh Chu-LaGraff

Abstract

Unlabeled primary immunoglobulin G (IgG) antibodies and its F(ab')2 and Fc fragments were attached to oxygen-plasma-cleaned glass substrates using either microcontact printing (MCP) or physical adsorption during bath application from dilute solutions. Fluorescently labeled secondary IgGs were then bound to surface-immobilized IgG, and the relative surface coverage was determined by measuring the fluorescence intensity. Results indicated that the surface coverage of IgG increased with increasing protein solution concentration for both MCP and bath-applied IgG and that a greater concentration of IgG was transferred to a glass substrate using MCP than during physisorption during bath applications. Scanning force microscopy (SFM) showed that patterned MCP IgG monolayers were 5 nm in height, indicating that IgG molecules lie flat on the substrate. After incubation with a secondary IgG, the overall line thickness increased to around 15 nm, indicating that the secondary IgG was in a more vertical orientation with respect to the substrate. The surface roughness of these MCP patterned IgG bilayers as measured by SFM was observed to increase with increasing surface coverage. Physisorption of IgG to both unmodified patterned polydimethyls...Continue Reading

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Citations

Oct 18, 2012·Analytical Chemistry·Kazuyo ShigetaJohn A Rogers
Apr 15, 2014·Colloids and Surfaces. B, Biointerfaces·Maria Dąbkowska, Zbigniew Adamczyk
Jan 17, 2015·Sensors·Erik Reimhult, Fredrik Höök
Aug 5, 2008·Biosensors & Bioelectronics·Kevin A HeyriesChristophe A Marquette
Jul 25, 2013·Scientific Reports·Xiaohua XuMoses Rodriguez
Nov 13, 2007·Journal of Biomedical Materials Research. Part a·Adam W FeinbergAnthony B Brennan
Mar 7, 2008·Sensors·Dorothee GrieshaberErik Reimhult
Sep 15, 2017·Engineering in Life Sciences·Cindy Dirscherl, Sebastian Springer
Feb 29, 2008·Langmuir : the ACS Journal of Surfaces and Colloids·Jennifer O FoleyPaul Yager
Sep 7, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Katarzyna DerwinskaClaudia Preininger
Oct 16, 2020·Langmuir : the ACS Journal of Surfaces and Colloids·Didac MartíJuan Torras
Mar 23, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Naïs CoqWendy U Dittmer
Mar 16, 2007·Langmuir : the ACS Journal of Surfaces and Colloids·Kevin A HeyriesLoïc J Blum

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