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Settlement inhibiting method of tiny particle for binding reactive substance and the reagent

A micro-particle and reactive technology, which is applied in. Especially in the field of immunization that utilizes antigen-antibody reaction mainly in the field of clinical examination, it can solve the problem of not inhibiting micro-particles, and achieve the effect of inhibiting sedimentation.

Inactive Publication Date: 2008-09-17
ALFRESA PHARMA CORP
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] Inhibition of the settling of such tiny particles that bind reactive species has not been done before

Method used

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  • Settlement inhibiting method of tiny particle for binding reactive substance and the reagent
  • Settlement inhibiting method of tiny particle for binding reactive substance and the reagent
  • Settlement inhibiting method of tiny particle for binding reactive substance and the reagent

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0060] Embodiment 1: the preparation of colloidal gold solution

[0061] Add 2mL of 10% chloroauric acid solution to 1L of 95°C distilled water while stirring, add 10mL of 2% sodium citrate solution after 1 minute, stir for another 20 minutes, and then cool to 30°C. After cooling, adjust the pH to 7.1 with 0.1% calcium carbonate.

Embodiment 2

[0062] Example 2: Preparation of the first reagent for determination of serum cystatin C

[0063] Add 1.0-2.5% to the 0.5M Bis-Tris (pH6.7) solution containing 5% sodium chloride, 0.2% EDTA, 0.2% alkylphenyl disulfonic acid sodium salt, 0.35% polyoxyethylene lauryl ether The left and right polyethylene glycols are used as reaction accelerators to prepare the first reagent for measuring serum cystatin C.

Embodiment 3

[0064] Embodiment 3: the preparation of the colloidal gold reagent (second reagent) that binds anti-serum cystatin C antibody

[0065] Anti-cystatin C antibody (Dako Japan Co., Ltd.) was diluted with 10 mM HEPES (pH 7.1) containing 0.05% sodium azide to a concentration of 50 μg / ml. 100 mL of the anti-cystatin C antibody solution Add to about 1L of the colloidal gold solution prepared in Example 1, and stir for 2 hours in a refrigerated state. Then, add 110mL of 10mM HEPES containing 5.46% mannitol, 0.5% BSA and 0.05% sodium azide .1), stirred at 37°C for 90 minutes. Then, centrifuged at 8000 rpm for 40 minutes, removed the supernatant, and then added about 1L of 5mM solution containing 3% mannitol, 0.1% BSA and 0.05% sodium azide HEPES (pH7.5) (A solution) to disperse the colloidal gold bound to the antibody. Then, centrifuge at 8000 rpm for 40 minutes, remove the supernatant, and disperse the antibody-sensitized colloidal gold in the A solution, so that the total amount reach...

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Abstract

The invention provides a method for keeping the uniform concentration of the small particles for combining the reactive substance in dispersion liquid by preventing the small particles for combining the reactive substance from subsiding in dispersion liquid. The invention provides a settling prevention method of the small particles for combining the reactive substance. The method comprises the step of coexisting at least one of polyanion and salt thereof, glucan, cyclodextrin, polyethylene glycol and glycerine in the dispersion liquid of the small particles.

Description

technical field [0001] The present invention relates to a method for inhibiting sedimentation of microparticles to which reactive substances such as antibodies and antigens are bound (hereinafter referred to as microparticles to which reactive substances are bound) and reagents for immunoassays using the microparticles to which reactive substances are bound .In particular, to a method for inhibiting sedimentation of microparticles that bind reactive substances used in immunological assays utilizing antigen-antibody reactions mainly in the field of clinical examinations, and reagents for immunoassays using the microparticles that bind reactive substances . Background technique [0002] In recent years, for various examinations such as clinical examinations, from the viewpoint of automation and shortening the measurement time, methods for measuring substances in biological samples using immunoreactions have been widely used. Examples of such immunoassay methods include the RIA...

Claims

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Application Information

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IPC IPC(8): G01N33/53G01N33/531B01J13/00G01N33/543G01N33/553
Inventor 田中睦
Owner ALFRESA PHARMA CORP
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