SPR (surface plasma resonance) analyzing method for determining rutin and protein binding level

A technology of surface plasmon and analysis method, which is applied in the field of surface plasmon resonance analysis to determine the binding level of rutin and protein, and can solve the problems of rutin solubility reduction and other problems

Inactive Publication Date: 2017-08-18
PEKING UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

In particular, in order to simulate the physiological environment, the SPR analysis process often uses a buffer solution containing inorganic salts to prepare the sample solution, which further leads to the problem of reduced solubility of rutin
There is no method of using SPR method to study the interaction between rutin and protein in the existing patent method

Method used

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  • SPR (surface plasma resonance) analyzing method for determining rutin and protein binding level
  • SPR (surface plasma resonance) analyzing method for determining rutin and protein binding level
  • SPR (surface plasma resonance) analyzing method for determining rutin and protein binding level

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Experimental program
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Embodiment

[0044] Embodiment: The analysis method of the present invention is used to determine the equilibrium constant of the interaction between rutin and human serum albumin (HSA). Specific steps are as follows.

[0045] 1. Use a new sensor chip, which is a glass plate coated with a bare gold film. Install it into the SPR instrument, start the instrument, and maintain the temperature of the chip area at 37°C.

[0046] 2. Use 5mmol / L 3-mercaptopropionic acid aqueous solution to circulate for 30 minutes at a flow rate of 5 microliters / minute, and control the temperature of the chip area at 37°C. After completion, rinse the pipeline and chip surface with water.

[0047] 3. Dissolve HSA in acetic acid-sodium acetate buffer solutions with a sodium ion concentration of 10 mmol / L and a pH of 4.0, 4.5, 5.0, and 5.5, respectively, and circulate for 5 minutes at a flow rate of 5 μl / min. The pH of the solution with the largest signal change at the end of the circulation is 5.5, so the buffer ...

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Abstract

The invention discloses an SPR (surface plasma resonance) analyzing method for determining rutin and protein binding level. An analyzing object is a rutin and protein interaction system, and an analyzing target is to solve an equilibrium constant of the interaction reaction. The method has the advantages that the rutin is dissolved by pyridine, and is diluted by a buffer solution containing the pyridine, so that the problem of insufficient dissolvability of the rutin in a water solution is solved, and an enough strong SPR signal is produced; when the diluted rutin solution is prepared in the analyzing process, the pyridine diluting solutions containing same components but not containing the rutin are correspondingly prepared, and are used for determining the influence of the pyridine to the SPR signal, so as to accurately obtain an interaction signal generated by the rutin.

Description

technical field [0001] The invention relates to an application method of surface plasmon resonance technology in analytical chemistry, in particular to a surface plasmon resonance analysis method for measuring the interaction and binding level of rutin and protein. Background technique [0002] Surface plasmon resonance (SPR) technology is a widely used method in the field of analysis of intermolecular interactions. The basic principle is to irradiate the SPR sensor chip with incident light, so that the evanescent wave generated by the incident light resonates with the surface plasmon wave of the metal on the surface of the sensor chip, which affects the nature of the outgoing light; after that, an interaction reaction occurs on the surface of the sensor chip , causing changes in the resonance conditions, leading to changes in the properties of the outgoing light, and monitoring it can realize the detection of the interaction reaction. In the general SPR analysis process, a...

Claims

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Application Information

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IPC IPC(8): G01N21/552G01N21/59G01N1/38
CPCG01N21/554G01N1/38G01N21/59G01N2001/383G01N2021/5903
Inventor 刘虎威白玉张一丁
Owner PEKING UNIV
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