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Affinity particle and affinity separation method

A separation method and affinity technology, applied in the field of affinity particles and affinity separation, can solve the problems of difficult separation of affinity particles, low capture efficiency, carrier disintegration, etc., to prevent non-specific adsorption, high separation selectivity, improve The effect of purification efficiency

Inactive Publication Date: 2007-05-02
SHISEIDO CO LTD +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are fatal problems shown in the following (1)-(3) in column separation
[0003] (1) In order to obtain the target substance, various columns must be used, and the purification efficiency is poor
[0004] (2) Confirmation experiments must be carried out to confirm whether the target substance is contained in the separated components, so purification takes more time
[0005] (3) The loss during purification is large, requiring a large amount of samples
[0009] (2) The capture efficiency is low and a large amount of liquid sample is required
[0012] (2) Light specific gravity, difficult to separate affinity particles
[0013] (3) Disintegration of the carrier is easy to occur, and the durability is low
[0014] On the other hand, since inorganic particles adsorb more substances than organic particles, those skilled in the art did not think of using inorganic particles as affinity particles

Method used

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Examples

Experimental program
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Embodiment

[0197] The present invention will be further described in detail below according to the examples. The present invention is not limited by these Examples. Phosphorylcholine groups introduced on the particle surface can be confirmed and quantified by FT-IR and elemental analysis.

[0198] "Synthesis Example 1"

[0199] "Aldehyde Compounds Containing a Phosphorylcholine Group"

[0200] Dissolve 6.29 g of 1-α-glycerophosphorylcholine in 210 ml of distilled water, and cool in an ice-water bath. Add 10.23 g of sodium periodate, and stir for 5 hours. The reaction solution was concentrated under reduced pressure, dried under reduced pressure, and the target compound was extracted with methanol. The following compound (6) shows its structure.

[0201] The 1H NMR spectrum of the compound of formula (6) in heavy water is shown in Figure 2. The compound of formula (6) is in equilibrium with formula (9) in water, so the actual spectrum reflects both formula (6) and formula (9).

[0...

reference example 1

[0231] The 13C-CPMAS spectrum and 13C-PSTMAS spectrum of the PC particles (A) of Reference Example 1 are shown in FIG. 5 . PSTMAS spectroscopy is a method for selectively obtaining the spectrum of free-moving molecular chains, and is widely used for the analysis of modified chains on particle surfaces. In Fig. 5, a spectrum of carbon derived from a phosphorylcholine group was observed at 54.2 ppm.

[0232] In the 31P-CPMAS spectrum of the PC particles (A) of Reference Example 1 shown in Figure 6, the NaH measured as the object 2 PO 4 Peaks were detected at almost the same chemical shift values, thereby confirming the presence of phosphate groups. From the above results, it can be considered that phosphorylcholine groups can be introduced into the surface of the carrier silica gel.

[0233] In FIG. 5 , spectra derived from the carbon of the spacer propyl group were observed around 9 ppm and 23 ppm, and spectra derived from the ethyl group in phosphorylcholine were observed a...

Embodiment 1

[0237] "Example 1"

[0238] "Affinity particles (Af particles (A)) having phosphorylcholine groups and amino groups in the form of covalent bonds on the surface of inorganic particles"

[0239] Take 87.9 μl of methanol solution containing 45 μmol of the compound of formula (10) prepared in Synthesis Example 3 and 50 μl of methanol solution containing 5 μmol of 3-aminopropyltrimethoxysilane, add 47.5ml of methanol, 2.5ml of distilled water, and then add 5g of the average Particle size is 1.5μm, specific surface area is 6m 2 / g of silica gel. The particle dispersion solution was refluxed overnight at 80° C. for coupling. After reflux, the affinity particles (hereinafter referred to as Af particles (A)) according to claim 1 were obtained by centrifugal washing with methanol. The P quantitative measurement of the Af particles (A) treated with the surface modifier of Synthesis Example 3 in the above procedure is shown in FIG. 4 . From this, particles with a PC introduction amou...

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Abstract

The present invention is affinity particles that are characterized by having phosphorylcholine groups represented by the following formula (1) covalently bonded onto the surface of inorganic powder and also by having ligands having specific affinity with a certain target substance covalently bonded or adsorbed onto the surface of inorganic powder. The object of the present invention is to provide an affinity separation method that uses affinity particles utilizing inexpensive inorganic particles and is capable of separating the target substance easily and with high accuracy.

Description

technical field [0001] The present invention relates to affinity particles and affinity separation methods. More specifically, it relates to an affinity separation method using affinity particles using inorganic particles and capable of separating target substances with high precision and easily. It is also extremely useful for various separation, purification, and detection methods such as immunoprecipitation and latex agglutination, which can detect target substances with high precision and easily. Background technique [0002] Conventionally, separation and purification of biological substances have been performed by column chromatography. However, there are fatal problems as shown in the following (1)-(3) in column separation. [0003] (1) In order to obtain the target substance, various columns must be used, and the purification efficiency is poor. [0004] (2) Confirmation experiments must be carried out to confirm whether the target substance is contained in the se...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01J20/22C07F9/09B01J20/286
Inventor 石原一彦前野克行宫泽和之石洼章
Owner SHISEIDO CO LTD
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