Name: In Focus - Surface plasmon resonance V6
Uploaded: 2023-09-20T00:00:00.0000000
Duration: T00H03M11S
Description: In Focus - Surface plasmon resonance V6

Name: In Focus - Surface plasmon resonance V6

Description: In Focus - Surface plasmon resonance V6

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Upload Date: 2023-09-20T00:00:00.0000000

Transcript: Language: EN.
Segment:0 .
Surface plasmon resonance, or SPR, is a real-time, label free optical technique used to detect and quantify molecular interactions. SPR has important applications in drug discovery and development and has many advantages over endpoint analysis techniques, such as ELISA, which have made it an attractive tool for analyzing molecular interactions.
But how does this phenomenon work, and what are its applications? SPR occurs under conditions of total internal reflection when incident light is reflected off a glass sensor chip coated with a thin gold film. The presence of free electrons at the interface between gold and glass is required to generate surface plasmons. When total internal reflection occurs at a specific angle, some of the energy from the incident light is lost.
This occurs when the light interacts with delocalized electrons in the thin metal film, which causes a reduction in the reflected light intensity, creating surface plasmons. These surface plasmons are highly sensitive to any changes that occur within close proximity of the surface utilized in the biosensor setup. For biosensor setups, the gold side of the sensor chip is coated with an appropriate surface chemistry that allows molecules to be immobilized or captured and is in contact with microfluidics through which assay buffer flows.
When a solution containing an analyte flows over the sensor chip, the analytes may bind to the ligands on the chip, causing the refractive index to change at the sensor chip surface. When the refractive index changes, the resulting signal is recorded in resonance units. Resonance units are both proportional to the total mass of the bound material as well as the number of bound biomolecules.
To remove the analytes from the ligand solution, a dissociation step can be completed using assay buffer. Any remaining bound analytes can be removed using a regeneration solution, which encourages dissociation. The refractive index at the sensor chip surface is monitored throughout the assay, allowing researchers to observe analyte association and dissociation to the ligand. With data collected from SPR assays, researchers can determine the kinetic rate constants and binding affinity of a given analyte to a target ligand.
Additionally, SPR can be used to assess binding specificity, which allows researchers to determine which analyte in a solution can bind preferentially to a target ligand. This technique can also provide an insight into analyte concentration. SPR can be a lengthy process requiring multiple analyte dilutions. So when it comes to choosing the right tool for your research, be sure to consider this limitation and how you might be able to circumvent it.
One potential solution is Sartorius' Octet SF3 SPR platform that has the OneStep injection function. Requiring only a single analyte concentration and injection to determine kinetic rate constants and binding affinity, this technology allows users to simplify assay development, decrease time to results and save both reagents and money. To find out more about surface plasmon resonance, check out our In Focus with Sartorius at www.BioTechniques.com.

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