title: In situ microarray fabrication and analysis using a microfluidic flow cell array integrated with surface plasmon resonance microscopy.
abstract: Surface Plasmon Resonance Microscopy (SPRM) is a promising label-free analytical tool for the real-time study of biomolecule interactions in a microarray format. However, flow cell design and microarray fabrication have hindered throughput and limited applications of SPRM. Here we report the integration of a microfluidic flow cell array (MFCA) with SPRM enabling in situ microarray fabrication and multichannel analysis of biomolecule probe-target interactions. We demonstrate the use of the MFCA for delivery of sample solutions with continuous flow in 24 channels in parallel for rapid microarray creation and binding analysis while using SPRM for real-time monitoring of these processes. Label-free measurement of antibody-antibody interactions demonstrates the capabilities of the integrated MFCA-SPRM system and establishes the first steps of the development of a high-throughput, label-free immunogenicity assay. After in situ probe antibody immobilization, target antibody binding was monitored in real time in 24 channels simultaneously. The limit of detection for this particular antibody pair is 80 ng/mL which is approximately 6 times lower than the industry recommended immunogenicity assay detection limit. The integrated MFCA-SPRM system is a powerful and versatile combination for a range of array-based analyses, including biomarker screening and drug discovery.
authors.affiliation: Department of Chemistry, University of Utah, Salt Lake City, Utah 84112, USA.
authors.forename: Jianping
authors.initials: J
authors.lastname: Liu
authors.forename: Mark A
authors.initials: MA
authors.lastname: Eddings
authors.forename: Adam R
authors.initials: AR
authors.lastname: Miles
authors.forename: Rostislav
authors.initials: R
authors.lastname: Bukasov
authors.forename: Bruce K
authors.initials: BK
authors.lastname: Gale
authors.forename: Jennifer S
authors.initials: JS
authors.lastname: Shumaker-Parry
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis* / Q000276:immunology*; D055910:Antibodies, Immobilized; D019921:Biotinylation; D002138:Calibration; D004867:Equipment Design; D006041:Goats; D006801:Humans; D007074:Immunoglobulin G / Q000032:analysis / Q000276:immunology; D046210:Microfluidic Analytical Techniques / Q000191:economics / Q000295:instrumentation* / Q000379:methods; D040081:Protein Array Analysis / Q000191:economics / Q000295:instrumentation* / Q000379:methods; D012680:Sensitivity and Specificity; D019809:Streptavidin; D020349:Surface Plasmon Resonance / Q000295:instrumentation / Q000379:methods*
chemical_list: D000906:Antibodies; D055910:Antibodies, Immobilized; D007074:Immunoglobulin G; D019809:Streptavidin