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Preparation method for immune nano magnetic glucan micro-sphere

A nano-magnetic and dextran technology, which is applied in the preparation methods of peptides, chemical instruments and methods, peptides, etc., can solve the problems of highly toxic cyanogen bromide, wide particle distribution, high price, etc. The effect of narrow particle size distribution and narrow particle size range

Active Publication Date: 2008-03-19
北京泽诚生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] But so far the commercialized products are mainly concentrated in foreign products. Although the performance is relatively good, the price is expensive, which discourages some research institutions and small and medium-sized enterprises. Moreover, the particle size of magnetic microspheres is mainly concentrated in the micron level, and the particle size is large. Some of the coated organic layers of magnetic microspheres, some of which are coated with some carbohydrates, are easy to fall off due to physical adsorption or connection through simple chemical bonds, and the distribution of particle diameters is wide and uneven (Steefan Miltenyi.: High Gradient Magnetic Cell Separation With MACS.JCytometry 11: 231-238 (1990), this is limited for some separations, and in the process of protein coating, cyanogen bromide is often used, but cyanogen bromide is highly toxic, and Ligands on substrates activated by cyanogen bromide often leak (Clonis YD, Small DAP. High-performance dye-ligand Chromatograph. ln Reactive in Protein and Enzyme Technology (M). London: Macmillan Press, 1 987.87-100)

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0040] Example 1: 30 ml of mixed solution of 2 g of ferric sulfate and 0.5 g of ferrous chloride, and add 0.1 g of nickel sulfate, then heated to 50 ° C, 2000 rev / min speed stirring at 4-20 ml / min speed 10 milliliters of 0.6 mol / L sodium hydroxide solution was added dropwise. After the sodium hydroxide was added dropwise, the temperature was raised to 80° C. and reacted for 1 hour; washed with deionized water for 4 times. Store in 20mmol / L phosphate buffer solution at 4°C.

Embodiment 2

[0041] Example 2: Add 30g of dextran 40 to 30ml of 0.1M acetate buffer solution to dissolve and mix well; add 9ml of 0.1mol / L sodium periodate solution, and react in the dark at 25°C for 1 hour, and the reaction ends Then dialyze with deionized water for 20 hours at 4°C; store the oxidized dextran in 30ml deionized water, and slowly drop 30ml of 1 Mixture of 50% activated dextran by volume and 50% unactivated dextran by volume, stirred at 500 rpm, reacted at 4°C for 48 hours, added 3ml of 4mg / ml sodium borohydride solution, mixed well, and kept at 4°C React for 2 hours. The above solution was dialyzed against 0.15M phosphate buffered saline at 4°C for 20 hours. Store in phosphate buffer at 4°C.

Embodiment 3

[0042] Example 3: Drain 2g nano-dextran magnetic microspheres, add 10ml of 0.8mol / L sodium hydroxide solution, mix well, add 1.5g epichlorohydrin, mix well, react at 25°C for 8 hours, acetone 1. Rinse with deionized water for 5 times, dilute nano-magnetic dextran microspheres to 80mg / ml with 25ml phosphate buffer; add 0.04g of 0.5mg / ml protein solution, react at 40°C for 8 hours, and carry out on a magnetic plate Separated, washed 5 times with phosphate buffered saline, preserved in phosphate buffered saline, and stored at 4°C.

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PUM

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Abstract

The present invention relates to a preparation method of immune nano-magnetic dextran microspheres, which belongs to the technical field of biological separation. In the process steps, nano-size magnetic iron oxide particles are prepared by the method of coprecipitation; a solution is prepared with ferrous ion, iron ion and nickel ion according to the molar ratio of 1:(1.0 to 4.0):(0.2 to 0.8); glucan enwraps the magnetic iron oxide particles; the proteins of the nano-magnetic dextran microspheres are linked. The present invention has the advantages of realization of the narrow diameter of particles, good reproducibility, low price and natural activity of the immune nano-magnetic dextran microspheres.

Description

technical field [0001] The invention belongs to the technical field of bioseparation, and in particular provides a convenient method for biomagnetic separation, which is applicable to various biological separations. Background technique [0002] The invention relates to a preparation method of dextran nano magnetic microspheres. The magnetic microspheres have a particle diameter of about 50-100 nm and are mainly used in nucleic acid, protein, cell and immunological testing. [0003] Dextran, also known as dextran, is a type of polysaccharide with a linear main chain, mainly connected with 1,6-α-D-pyranoside, itself and its degradation products in the body ( CO2, H2O) are non-toxic to the human body, and have been used clinically as a blood volume expander for more than 50 years. Dextran molecules can exist stably in the environment of weak acid or weak base (JANES KA, CALVO P, ALONSO MJ, Pollusaccharide collagen particles as delivery systems for macromolecules[J]. Adv Drug ...

Claims

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Application Information

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IPC IPC(8): C07K1/04C07K17/10
Inventor 陈海生张华英陈宝俊刘金静刘亚宁郭丽峰
Owner 北京泽诚生物技术有限公司
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