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Streptavidin-bonded magnetic particles and manufacturing method for same

A streptavidin, magnetic particle technology, applied in biochemical equipment and methods, material testing products, instruments, etc., can solve the problems of reduced washing efficiency, reduced sensitivity, and increased turbidity.

Inactive Publication Date: 2013-12-11
KYOWA MEDEX CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0007] (1) Since the magnetic particles will settle under static conditions, it is necessary to disperse them during use, but if the amount of particles is large, it will take time and labor to disperse
[0008] (2) If the amount of particles is large, the volume will increase when the magnet is used to bring it closer to one side of the container, and the efficiency of washing the reaction liquid enclosed in the inside during B / F separation and washing will decrease.
[0009] (3) When detecting the component to be measured, if the amount of magnetic particles is large, the turbidity will increase due to the coloring of the magnetic particles themselves. For example, in the detection by chemiluminescence or fluorescence, the optical shielding will cause the Reduced sensitivity
However, there are problems that if the particle size is reduced, the time for collecting particles by magnets will be greatly prolonged, or the particles will become easy to flow during the spraying of the cleaning solution in the washing process and the suction operation, etc.
For example, Patent Document 2 describes a method of chemically forming a porous layer on the outer layer of magnetic particles. In this method, in the immune reaction between antigen and antibody or in the hybridization between DNA or DNA and RNA, the unit surface area The binding energy is improved, but the reaction efficiency in the hole is poor, and it is difficult to obtain the expected performance

Method used

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  • Streptavidin-bonded magnetic particles and manufacturing method for same
  • Streptavidin-bonded magnetic particles and manufacturing method for same
  • Streptavidin-bonded magnetic particles and manufacturing method for same

Examples

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Embodiment 1

[0199] (1) Manufacture of streptavidin-conjugated magnetic particles

[0200] As the magnetic particles, amino-type Estapor magnetic particles EM2-100 / 40 (manufactured by Merck) were used. The magnetic particle is composed of a core-shell structure with a particle size of 1.62 μm, the inner core part contains a magnetic substance with a total mass ratio of 41.2%, and the shell part composed of polystyrene is chemically modified with amino groups at a rate of 97 μeq / g. particle. 2 mg of the magnetic particles were collected, and a pH 5.5, 10 mmol / L acetate buffer (hereinafter referred to as dispersion A) containing 1.0% of Trimethylstearylammonium Chloride (manufactured by Tokyo Chemical Industry Co., Ltd.) was added. 4mL to disperse it. Next, a powerful magnet is placed on the side of the container to collect the magnetic particles and remove the dispersion A by suction (hereinafter, the series of operations of adding the dispersion, collecting the magnetic particles, and su...

Embodiment 2

[0212] Analysis of the Crosslinked Structure of Streptavidin on Magnetic Particles

[0213] For the biotin binding capacity obtained by the same method as that of Example 1 is 2.61 pmol / mm 2 、4.95pmol / mm 2 、6.76pmol / mm 2 Each streptavidin-bound magnetic particle was washed 10 times with 4 mL of PBS, replaced with 1% SDS / PBS, and incubated at 60°C for 1 hour. Next, the magnetic particles were collected by a magnet, and the protein solution in the supernatant was collected and analyzed by SDS-PAGE. The same operation was also performed on commercially available streptavidin-conjugated magnetic particles. The results of SDS-PAGE are shown in figure 1 .

[0214] Such as figure 1 As shown, in streptavidin and commercially available streptavidin-bound magnetic particles, only monomers constituting streptavidin can be seen, but in the streptavidin-bound magnetic particles of the present invention, In addition to monomers, dimers, trimers, tetramers and higher order bands can a...

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Abstract

Provided are streptavidin-bonded magnetic particles having a high biotin binding capability, and a manufacturing method for the same. Streptavidin-bonded magnetic particles are characterized by having a structure in which streptavidins are cross-linked on magnetic particles. A manufacturing method for streptavidin-bonded magnetic particles including the following steps. (1) A step for preparing a suspension containing magnetic particles having amino groups on the surface thereof; and (2) a step for adding glutaraldehyde to the suspension prepared in step (1), in the presence of streptavidin, and reacting the magnetic particles with the streptavidin and glutaraldehyde. These streptavidin-bonded magnetic particles, and streptavidin-bonded magnetic particles manufactured by means of this manufacturing method can be used in clinical diagnosis.

Description

technical field [0001] The present invention relates to streptavidin-bound magnetic particles (streptavidin-coupled magnetic particles) and methods for producing the same, and protein-bound magnetic particles (protein-coupled magnetic particles) produced using the streptavidin-coupled magnetic particles A method for producing the same, a method for measuring a component to be measured, and a reagent for measuring a component to be measured. Background technique [0002] In diagnostic medicines, magnetic particles are often used as solid phase carriers in order to detect test substances such as hormones, cancer markers, and infectious disease markers. In this assay system, antibodies or antigens (primary probes), etc. are bound to magnetic particles, and after binding to the substance to be measured in the specimen, they are then passed through a fluorescent substance or chemiluminescent substrate, enzyme, etc. labeled The secondary probe binds to qualitatively or quantitati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/53G01N33/543G01N33/553
CPCG01N33/531G01N33/54326G01N33/5434G01N33/53G01N33/543G01N33/553C12Q1/6816G01N33/54346G01N33/566
Inventor 荒井信之松冈泰弘守田和树
Owner KYOWA MEDEX CO LTD
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