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Contrary to the fact that SPR currently plays a very important role in the fields of life science, materials, and pharmaceuticals, the reality is that there are few opportunities to learn the theory and experiment of SPR at universities and even graduate schools. icluebio is operating an SPR education center so that practitioners and researchers in the field can understand and use SPR. If you need training, please apply for regular training by filling out the training application form below. You can learn simple SPR theory and device principles through regular free training, and you can discuss with experts whether the experiment you need to perform is possible. Customers who have already purchased it can learn new upgraded features.
SPR refers to a phenomenon in which the reflected light disappears at a specific incident angle when light is incident on the side of a prism on which a gold film (about 50 nm) is deposited, and the angle at this time is called the SPR angle. When biomolecules combine on the gold film, the refractive index of the gold film surface changes, and the SPR angle moves according to the refractive index change. The SPR system characterizes intermolecular bonds by acquiring sensorgrams that record changes in the SPR angle in real time.
What is the SPR and how can intermolecular bonding be measured?
3 steps for SPR measurement
Ligand ImmobilizationThe ligand immobilization step is to immobilize the ligand on the surface of the sensor chip. There are a wide variety of materials used as ligands, such as antibodies, proteins, nucleic acids, and peptides. Depending on the characteristics of the ligand material, the surface functional group (COOH, NTA), and the immobilization method.
Analyte binding is divided into an association step in which an analyte solution is injected and bound to the ligand and the graph rises in real-time, and a dissociation step in which some analytes attached to the ligand are dropped by injecting only a running buffer. Then, if the remaining analyte attached to the ligand is removed by the regeneration solution, the same sensor chip can continue to experiment with different concentrations or different types of analytes.
Kinetic parameters can be calculated through sensorgrams in which association and dissociation sections are observed by flowing analytes of various concentrations. You can usually find the association rate constant (ka) and dissociation rate constant (kd) through a 1:1 binding model. Using this, the equilibrium dissociation constant (KD, Equilibrium dissociation constant) can also be calculated.
What analysis can be done using the SPR system?
Yes/No: Check the binding. By simply flowing the analyte (B) onto the sensor chip on which the ligand (A) is immobilized, the binding is yes or no can be checked.
Screening: Quickly discover candidates from your library with one representative concentration measurement.
Concentration: After creating a standard quantitation curve, the concentration of the analyte can be determined.
Affinity: You can evaluate the equilibrium constant after acquiring the sensorgram up to the equilibrium stage and creating the Langmuir isothermal curve.
Kinetics: You can evaluate the rate constants ka and kd through curve fitting by acquiring the association/dissociation section of the sensorgram.
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