Surface plasmon resonance imaging measurements of antibody arrays for the multiplexed detection of low molecular weight protein biomarkers

Hye Jin Lee, Dobrin Nedelkov, Robert M. Corn

Research output: Contribution to journalArticlepeer-review

165 Scopus citations

Abstract

This paper describes a simple methodology for the creation of high-density multiplexed antibody arrays on gold surfaces that can be used to detect low molecular weight protein biomarkers with surface plasmon resonance imaging (SPRI). A one-step carbonyldiimidazole (CDI) surface reaction was utilized to attach antibodies onto alkanethiol-modified gold surfaces and characterized with polarization modulation FT-IR reflection absorption spectroscopy. The CDI chemistry was then employed to create an antibody microarray with array element sizes varying from 750 μm down to 200 μm. As a demonstration, a three-component antibody array was employed to detect two clinically important protein biomarkers, β2-microglobulin (11.8 kDa) and cystatin C (13.4 kDa). SPRI measurements could simultaneously detect both of these small unlabeled proteins with no cross talk at solution concentrations from 300 nM down to 1 nM. In addition, the adsorption strengths of these biomarkers onto an antibody array were measured with SPRI and compared to those obtained from the kinetic analysis of single-channel angle shift SPR measurements.

Original languageEnglish
Pages (from-to)6504-6510
Number of pages7
JournalAnalytical Chemistry
Volume78
Issue number18
DOIs
StatePublished - 15 Sep 2006

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