Method for measuring relative absorption constants of dye, antibody, medicament and medicament prosoma molecules on polypeptide molecules

An adsorption constant, molecular technology, used in the field of biomedicine

Inactive Publication Date: 2011-05-18
THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, some gaps inevitably exist between computer simulations and experimental studies.

Method used

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  • Method for measuring relative absorption constants of dye, antibody, medicament and medicament prosoma molecules on polypeptide molecules
  • Method for measuring relative absorption constants of dye, antibody, medicament and medicament prosoma molecules on polypeptide molecules
  • Method for measuring relative absorption constants of dye, antibody, medicament and medicament prosoma molecules on polypeptide molecules

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] Example 1 Determining the Relative Adsorption Constant of Dye Molecules on Polypeptide Based on Scanning Tunneling Microscope

[0049] 1. The chemical structure of the substance used

[0050] The chemical structures of polypeptide molecules (5Ala, 8Phe and 8His), pyridine molecules (4Bpy) and dye molecules sulfophthalocyanine are as follows:

[0051]

[0052] Pentapolyalanine (5Ala)

[0053]

[0054] Octaphenylalanine (8Phe)

[0055]

[0056] Octahistidine (8His)

[0057]

[0058] 4'4-bipyridine (4'4-bipyridyl, 4Bpy)

[0059]

[0060] Phthalocyanine-tetrasulfonic

[0061] 2. Measurement method

[0062] First, mix the polypeptide molecules (5Ala, 8Phe and 8His) and the pyridine marker molecule 4Bpy in the aqueous solution, and ultrasonicate for 10 minutes. After mixing thoroughly, take out 15 microliters of the solution, drop it on the surface of the newly cleaved graphite, and let it stand for 10 minutes. After the mixed molecular system is assemb...

Embodiment 2

[0068] Example 2 Determination of Relative Adsorption Constant of Drug Molecules on Polypeptides Based on Scanning Tunneling Microscopy

[0069] 1. The chemical structure of the substance used

[0070] The chemical structures of the polypeptide molecule pentapolyalanine (5Ala), the pyridine marker molecule (4Bpy) and the drug molecule Congo red (Congo red) are as follows:

[0071]

[0072] Pentapolyalanine (5Ala)

[0073]

[0074] 4'4-bipyridine (4'4-bipyridyl, 4Bpy)

[0075]

[0076] Congo red

[0077] 2, assay method is identical with embodiment 1

[0078] High-resolution STM images of Congo red adsorbed in the 5Ala and 4Bpy two-component assembly system were obtained by scanning tunneling microscopy (such as Figure 5 shown). By counting the amount of adsorption of Congo red on polypeptide and 4Bpy, the basis for the relative adsorption capacity of Congo red on polypeptide and marker molecule 4Bpy is provided (such as Figure 6 shown).

Embodiment 3

[0079] Example 3 Determination of Relative Adsorption Constant of Drug Prodrug Molecule on Polypeptide Based on Scanning Tunneling Microscopy

[0080] 1. The chemical structure of the substance used

[0081] The chemical structures of the polypeptide molecule pentapolyalanine (5Ala), the pyridine marker molecule (4Bpy) and the prodrug molecule Thioflavin (Thioflavin T, ThT) are as follows:

[0082]

[0083] Pentapolyalanine (5Ala)

[0084]

[0085] 4'4-bipyridine (4'4-bipyridyl, 4Bpy)

[0086]

[0087] Thioflavin (Thioflavin T, ThT)

[0088] 2. Measurement method

[0089] The assay method is the same as in Example 1, and a scanning tunneling microscope is used to obtain a high-resolution STM image (such as Figure 7 shown). By counting the adsorption quantity of ThT on the polypeptide and 4Bpy, the basis for the relative adsorption capacity of ThT on the polypeptide and the marker molecule 4Bpy is provided. Since ThT has no adsorption on 4Bpy, the relative ads...

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Abstract

The invention relates to a method for measuring the relative absorption constants of molecules to be measured, such as dye, an antibody, a medicament, a medicament prosoma, and the like on polypeptide molecules by utilizing a STM (Scanning Tunneling Microscopy) technology. The method comprises the following steps of: acquiring an STM image of the self-assembly structure of the polypeptide molecules or acquiring an STM image of the co-assembly structure of polypeptide-labeled molecules through co-absorption with labeled molecules, adding molecules to be measured, such as the dye, the antibody, the medicament, the medicament prosoma, and the like to acquire the combination sites and the absorption quantity of the molecules to be measured on polypeptide and search the interaction of the polypeptide and the molecules to be measured on a molecule level; and measuring the relative absorption constants of the molecules to be measured on the polypeptide. The method provided by the invention is very significant to the analysis of the interaction mode and the combination essence of the target protein and the medicament.

Description

technical field [0001] The invention belongs to the technical field of biomedicine. Specifically, the invention relates to a method for measuring the relative adsorption constants of molecules such as dyes, antibodies, drugs and drug prodrugs on polypeptides by using scanning tunneling microscopy (STM) . Background technique [0002] In the research of drug design and development, it is extremely important to analyze the interaction mode and binding nature of the target protein and the drug. Generally, high-throughput protein crystallization, high-resolution X-ray crystallography, and two-dimensional nuclear magnetic resonance can be used to analyze Conduct research (see Daniel Picot, Patrick J. Lollet al., Nature 1994, 367, 243. and Aneta T. Petkova, Yoshitaka Ishii et al., Proc. Natl. Acad. Sci. U.S.A. 2002, 99, 16742.) . [0003] Now X-ray crystallography technology is very mature, but due to the difficult crystallization of some important protein (such as membrane prot...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N13/00
CPCG01N33/52
Inventor 毛晓波杨延莲王琛
Owner THE NAT CENT FOR NANOSCI & TECH NCNST OF CHINA
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