Solution for dissociating vitamin d from vitamin d-binding protein, associated detection method and use

A vitamin and solution technology, applied in biological testing, material inspection products, etc., can solve problems such as insufficient dissociation

Active Publication Date: 2018-11-23
BIOMERIEUX SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0015] Binding of 25-hydroxyvitamin D, or more generally vitamin D and its metabolites, to vitamin D-binding protein (DBP) complicates determination of its composition
Applicants have used this solution, but found that the dissociation obtained was still insufficient

Method used

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  • Solution for dissociating vitamin d from vitamin d-binding protein, associated detection method and use
  • Solution for dissociating vitamin d from vitamin d-binding protein, associated detection method and use
  • Solution for dissociating vitamin d from vitamin d-binding protein, associated detection method and use

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0097] Example 1 - Advantages of dissociation using a mixture of methanol and PFCA compared to PFCA alone

[0098] Preparation of dissociation solution

[0099] Contrast dissociation solution: the components (5mM disodium hydrogen phosphate (Na 2 HPO 4 ), 1.5mM potassium dihydrogen phosphate (KH 2 PO 4 ), 131 mM NaCl) and 0.75% perfluorohexanoic acid were dissolved in deionized water by stirring for about 30 minutes (min). The pH was adjusted to 7.2 using 6N NaOH.

[0100] Solution of the present invention: will be used to prepare the component of PBS damping fluid (5mM disodium hydrogen phosphate (Na 2 HPO 4 ), 1.5mM potassium dihydrogen phosphate (KH 2 PO 4 ), 131 mM NaCl), 0.75% perfluorohexanoic acid and 5% methanol were dissolved in deionized water by stirring for about 30 minutes (min). The pH was adjusted to 7.2 using 6N NaOH.

[0101] Method for Quantification of Total 25-OH Vitamin D

[0102] use Immunoassay machine (from bioMérieux) was used for immun...

Embodiment 2

[0114] Example 2 - Comparing different alcohols

[0115] The dissociation solution used in this example was prepared using the method explained for Example 1 in PBS buffer at pH 7.2. The nature and concentration of fluoroalkyl surfactants and alcohols were varied and are listed in Tables 2 and 3.

[0116] In addition, the method is as in Example 1.

[0117] Table 2: Use of PFCA

[0118] Contrast dissociation solution: PBS+1% perfluorohexanoic acid , no alcohol

[0119]

[0120] Dissociation solution of the present invention: PBS+1% perfluorohexanoic acid +5% Methanol

[0121]

[0122] Dissociation solution of the present invention: PBS+1% perfluorohexanoic acid +5% ethanol

[0123]

[0124] Dissociation solution of the present invention: PBS+1% perfluorohexanoic acid +5% isopropanol

[0125]

[0126] It can be seen that the addition of alcohol (whatever the alcohol) leads to improved dissociation (decrease in signal ratio in %).

[0127] Table 3: Use ...

Embodiment 3

[0135] The influence of embodiment 3-perfluoroalkanoic acid concentration

[0136] The dissociation solution used in this heat exchanger was prepared using the method explained in Example 1 in PBS buffer at pH 7.2.

[0137] In addition, the method is as in Example 1.

[0138] Table 4

[0139]

[0140]

[0141] It can be seen that in all cases the dissociation increases in the presence of alcohol. It can also be seen that by increasing the concentration of perfluoroalkanoic acid, the dissociation is further increased.

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Abstract

The present invention relates to the use of at least one fluoroalkyl surfactant and of at least one alcohol comprising from 1 to 4 carbon atoms, for dissociating vitamin D, and / or a vitamin D metabolite, from vitamin D-binding protein, and also to a solution comprising at least one fluoroalkyl surfactant and at least one alcohol comprising from 1 to 4 carbon atoms. The subject matter of the invention is also a method for detecting and quantifying, in vitro, vitamin D and / or at least one vitamin D metabolite, in a biological sample, comprising the use of at least one fluoroalkyl surfactant andof at least one alcohol comprising from 1 to 4 carbon atoms, so as to dissociate vitamin D and / or a metabolite or metabolites thereof to be detected, from vitamin D-binding protein.

Description

[0001] This application is a divisional application with the application number 2014800511768, the application date is September 16, 2014, and the title of the invention is "a solution for dissociating vitamin D from vitamin D binding protein, related detection methods and uses". technical field [0002] The invention relates to the technical field of vitamin D detection. More specifically, the present invention provides the use of a combination of a fluoroalkyl surfactant and an alcohol for the release of vitamin D and / or one of its metabolites from a vitamin D binding protein, a solution comprising such a combination, using such In vitro method for combined detection / quantification of vitamin D and / or at least one metabolite of vitamin D, and kits for detection / quantification using such combination by immunoassay. Background technique [0003] Vitamin D is an important substance involved in many biological processes in humans and animals. Among them, vitamin D, known to b...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/82
CPCG01N33/82
Inventor 瓦莱丽·默尼耶伊曼纽尔·莫罗
Owner BIOMERIEUX SA
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