Encyclopedia of Experiments: Biological Techniques
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Transcript
To perform surface plasmon resonance, or SPR, for studying the biomolecular interactions, begin with a sensor chip positioned over an SPR detector in a flow cell. This metal-coated sensor chip has a layer of hydrogel material deposited on the chip surface, making it better suited for protein immobilization.
Overlay the chip with viral peptides, which interact with the hydrogel and immobilize on the surface. Wash with an alkaline quenching solution which prevents non-specific protein interactions.
Direct polarized light onto the chip. At the resonance angle, the metal electrons absorb this light and resonate, reducing the reflected light intensity at the corresponding reflected angle.
The photodetector collects this reflected light, and the processor converts this signal to baseline, generating a sensorgram — a graph representing the sensor response against time.
Flow a solution of antiviral agent into the flow cell. During the flow, the antiviral agents interact with the immobilized viral peptides on the chip. This association changes the refractive index, which alters the angle and intensity of reflected light, ultimately, resulting in a sensor response.
Add a washing buffer of high ionic strength that disrupts the interaction between the two molecules, leading to their dissociation and decreasing the sensor response.
Analyze the shape of the sensorgram, and assess the association and dissociation of the two molecules crucial in biomolecular interactions.
