Unlock instant, AI-driven research and patent intelligence for your innovation.

Determination of binding constants by means of equilibrium shifting

A technology of binding constants and dissociation constants, applied in measuring devices, biological tests, instruments, etc., can solve problems such as uncertain interactions

Active Publication Date: 2017-08-29
3B药品有限责任公司
View PDF3 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The disadvantage of this method is that the interaction between substances exhibiting very high or low affinity for the membrane and the target cannot be determined

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Determination of binding constants by means of equilibrium shifting
  • Determination of binding constants by means of equilibrium shifting
  • Determination of binding constants by means of equilibrium shifting

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0066] A) Experimental design

[0067] A particularly preferred embodiment is a microtiter plate used in the above method, in which 5 columns have 7 chambers, so that there are 25 samples with immobilized target and 10 samples without immobilized target as a control . Human serum albumin (HSA) was used as the immobilized target (Target 1). HSA was immobilized on commercial agarose beads (Mini-Leak beads medium, Kem-En-Tec Nordic A / S) according to a standard method, and human plasma was used as a soluble target. The substance to be tested is a peptide. Five concentrations of immobilized HSA were used for each of the five plasma concentrations. For this, the chambers of the microtiter plate are filled as follows:

[0068] Column I of the microtiter plate (use rows 1 to 7):

[0069]

[0070]

[0071] Column II of the microtiter plate (use rows 1 to 7):

[0072]

[0073]

[0074] Column III of the microtiter plate (use rows 1 to 7):

[0075]

[0076] Column I...

Embodiment 2

[0097] A) Experimental design

[0098] A particularly preferred embodiment is a microtiter plate used in the above method, in which 5 columns have 7 chambers, so that there are 25 samples with immobilized target and 10 samples without immobilized target as a control . Human serum albumin (HSA) was used as the immobilized target (Target 1). HSA was immobilized on commercial agarose beads (Mini-Leak beads medium, Kem-En-Tec Nordic A / S) according to several standard methods and human plasma was used as soluble target. The substance to be tested is liraglutide. Five concentrations of immobilized HSA were used for each of the five plasma concentrations. For this, the chambers of the microtiter plate are filled as follows:

[0099] Column I of the microtiter plate (use rows 1 to 7):

[0100]

[0101] Column II of the microtiter plate (use rows 1 to 7):

[0102]

[0103]

[0104] Column III of the microtiter plate (use rows 1 to 7):

[0105]

[0106]

[0107] Co...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention relates to a method for determining binding constants between a substance or a substance mixture and a target, and to a kit for carrying out the method according to the invention. The basic function principle of the method according to the invention, is the determination of the binding constants of the substance for the targets by shifting the binding equilibrium. In this context, the concentrations of the two targets, namely of the immobilised and of the dissolved target, are varied and the affinities with the targets are determined by the shifting of the binding equilibria in the individual batches. In this context, at least four samples are used, wherein, according to the invention, the first and the second sample of the substance comprising different substance amounts of the immobilized target and the same substance amount of the dissolved target are incubated, and the third and the fourth sample of the substance comprising the same substance amounts of the immobilized target as the first and the second sample of the substance and comprising the same substance amount of the soluble target are incubated, wherein the substance amount of the soluble target in sample containers three and four is different to the substance amount of the soluble target in the sample containers one and two, and the sample containers contain the same volume of liquid phase during incubation, consisting of a buffer, dissolved target and substance sample.

Description

technical field [0001] The invention relates to a method for determining the binding constant between a substance or substance mixture and a target according to claim 1 and to a kit according to claim 16 . Background technique [0002] The determination of interactions and binding constants between substances or substance mixtures and target substances plays an important role in the pharmaceutical industry, especially in drug research and development. Both fields deal with the interaction between substances or mixtures of substances and living things. Here, the binding properties of the individual substances must be quantified with binding parameters and binding constants. [0003] A method for determining the binding constant is known from DE 102010018 965 A1. In the method, the protein is immobilized on a plurality of solids, eg, contacted with an aqueous solution of the substance to be tested, and incubated for a period of time sufficient for the dissolved substance to ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): G01N33/543G01N33/92
CPCG01N33/54306G01N33/54313G01N33/92G01N33/48G01N33/50
Inventor 亨涅克·波瑞斯
Owner 3B药品有限责任公司