Abstract
The BIAcore (biomolecular interaction analysis) analytical system consists of a detection unit, an autosampler, and a liquid delivery system, controlled by a computer. Karlsson et al. (1) have given a detailed description of the analytical system, the detection principle, and the theoretical background for binding measurements. The system combines a microfluidic unit in contact with a sensor for surface plasmon resonance (SPR) detection. Figure 1 shows the principle of SPR detection. The sensor chip consists of a glass slide mounted in a plastic frame. On one side of the glass, a thin film, approx 50 nm, of gold is deposited, and the dextran matrix is attached on top of this film. The sensor chip is inserted into the instrument with the dextran/gold side in contact with the flow cells. When the injected sample is passing through the flow cell, antigen binds to immobilized antibody in the dextran matrix. Light covering a span of angles of incidence falls on the glass side and is reflected into a 2D array detector where the intensity of the reflected light is measured. SPR occurs at a certain angle and is seen as a minimum in reflected light intensity. When the refractive index close to the gold film is changed, for example, when immobilized antibody binds antigen, the angle at which SPR occurs is changed. This change is proportional to the amount of bound protein, and is expressed as refractive units (RU) on the Y-axis of a sensorgram; 1000 RU corresponds to 1 ng protein/mm2 of the 100-nm thick dextran layer. Reproducibility of the system has been validated by Fagerstam et al. (2).
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References
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© 1996 Humana Press Inc.
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Johne, B. (1996). Epitope Mapping by Surface Plasmon Resonance in the BIAcore. In: Morris, G.E. (eds) Epitope Mapping Protocols. Methods in Molecular Biology™, vol 66. Humana Press. https://doi.org/10.1385/0-89603-375-9:67
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DOI: https://doi.org/10.1385/0-89603-375-9:67
Publisher Name: Humana Press
Print ISBN: 978-0-89603-375-7
Online ISBN: 978-1-59259-552-5
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