期刊
ANALYTICAL CHEMISTRY
卷 78, 期 18, 页码 6504-6510出版社
AMER CHEMICAL SOC
DOI: 10.1021/ac060881d
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资金
- NCRR NIH HHS [1R21 RR018475-01A2] Funding Source: Medline
- NIGMS NIH HHS [2R01 GM059622-04] Funding Source: Medline
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 mu m down to 200 mu m. As a demonstration, a three-component antibody array was employed to detect two clinically important protein biomarkers, beta(2)-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.
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