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Decontaminant-doped dry hydrogel particles, macromolecule concentration and specific activity improvement

A technology of macromolecules and detergents, used in solid adsorbent liquid separation, hydrolase, filtration separation, etc., can solve the problems of low local concentration, polarization or loss of active function, clogging and other problems

Active Publication Date: 2019-04-05
福建省集力生物技术有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The present invention overcomes the disadvantages of existing macromolecular concentration techniques such as user unfriendliness, slowness, clogging, inefficient local concentration, polarization or loss of active function

Method used

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  • Decontaminant-doped dry hydrogel particles, macromolecule concentration and specific activity improvement
  • Decontaminant-doped dry hydrogel particles, macromolecule concentration and specific activity improvement
  • Decontaminant-doped dry hydrogel particles, macromolecule concentration and specific activity improvement

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] Embodiment 1: preferred conditions

[0020] The type of hydrogel particles: the type of hydrogel particles is selected according to the ease of preparation, preparation cost, and the need for interaction with target macromolecules. Two types of hydrogel particles, polyacrylamide and polydextran, are preferred because of their relative ease and economy of preparation and inertness that do not interact with macromolecules; polyacrylic acid or polyacrylic acid hydrogel particles are preferred because In addition to the ease and economy of its preparation, it also has a huge water absorption per unit gel mass, although the impact of the negative charge it carries on the interaction with the target macromolecule on the concentration process and recovery rate needs to be carefully considered.

[0021] The type and concentration of doped detergent: according to the characteristics of the interaction between target macromolecules and hydrogel particles, select the doped deterge...

Embodiment 2

[0025] Example 2: Preparation of detergent-doped dry polyacrylamide hydrogel particles by single-phase polymerization

[0026] Components: 29 grams of acrylamide, 1 gram of N,N-methylenebisacrylamide, 100 milliliters of deionized water, 0.2 grams of ammonium persulfate, and 40 microliters of tetramethylethylenediamine.

[0027] Steps: Dissolve and mix the above-mentioned acrylamide, N,N-methylenebisacrylamide and ammonium persulfate solids in deionized water in a beaker, add tetramethylethylenediamine to start the polymerization reaction, and immediately pour into the bottom area In a larger container with an inert inner surface, let it stand for about 1 hour to complete the polymerization; chop the polymerized gel and dry it at 90°C; crush the dried block polymer in a pulverizer, and screen out 60 ~150 target particles, poured into deionized water to fully swell; wash twice with 250 ml deionized water, and then use 250 ml Tween-20 deionized with a preferred concentration of 0...

Embodiment 3

[0028] Example 3: Preparation of detergent-doped dry polyacrylamide particles by reverse-phase suspension polymerization

[0029] Water phase configuration: Mix 29 grams of acrylamide, 1 gram of N,N-methylenebisacrylamide, 100 milliliters of deionized water and 0.2 grams of ammonium persulfate. Oil phase configuration: Mix 200ml cyclohexane and 3ml Span-80.

[0030] Steps: configure the oil phase in a three-necked flask equipped with mechanical stirring and a spherical condenser, raise the temperature to 57°C, and stir at a speed of 240 rpm; configure the water phase in the beaker, add it to the flask and mix with the oil phase, During the stirring operation, the water phase is dispersed into the oil phase, and the temperature is kept at 57°C for 30 minutes; the temperature is raised to 67°C, and a large amount of reflux occurs after the system releases heat, and the heat is kept for 30 minutes after the heat release; After the reaction, remove cyclohexane by vacuum filtratio...

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Abstract

The invention belongs to the field of macromolecule preparation, relates to decontaminant-doped dry hydrogel particles and a method for realizing mass concentration of a macromolecular liquid sample and specific activity improvement of a protein liquid sample by utilizing the decontaminant-doped dry hydrogel particles, and overcomes the defects of low efficiency, active function loss and the likein the prior art. The decontaminant-doped dry hydrogel particles are suitable for treatment of various sizes of macromolecular liquid samples. The hydrogel particles sufficiently suspend and are hydrated in a decontaminant solution, unnecessary decontaminant solution is filtered off, and then the particles are dried to obtain the decontaminant-doped dry hydrogel particles. The dry particles with cross-linked network apertures smaller than target macromolecules are enabled to contact with the macromolecular liquid sample proportionally, the hydrogel particles after imbibition are subjected to filtering centrifugation or conical filter membrane drum centrifugation so as to remove surface layer- dehydrated hydrogel particles, and then macromolecular mass concentrated filter liquid or proteinspecific activity improved concentrated filter liquid is obtained.

Description

technical field [0001] The invention relates to a dry hydrogel particle mixed with detergent, which realizes the mass concentration of macromolecules with high multiple and high recovery rate and the improvement of protein specific activity. Background technique [0002] The macromolecules referred to in the present invention include polysaccharides, nucleic acids, proteins, and aggregates formed by their single or composite components, such as natural biological macromolecules or aggregates such as ribosomes, exosomes, inclusion bodies, chromosomes, organelles, and viruses , Also includes water-soluble synthetic polymers or colloids such as polyacrylamide, polyacrylic acid, polyacrylate, polyvinyl alcohol, polyethylene glycol, polyvinyl pyrrolidone, latex particles, colloidal gold, etc. When preparing, modifying or analyzing liquid samples of these macromolecules, they often require concentration because they are too low in concentration. Precipitation caused by manipulati...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/08B01D36/00B01J20/28
CPCB01D15/08B01D36/00B01J20/28047B01D61/00B01J20/24B01J20/267C07K1/34B01D15/34B01D36/045B01D63/16B01J20/261B01J20/28016C07K1/36C07K14/765C12N9/0065C12N9/16C12Y111/01007C12Y301/03001
Inventor 黄志坚罗旻吕常庆
Owner 福建省集力生物技术有限公司
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