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Production method for protein molecule imprinting integral column

A technology of molecular imprinting and monolithic columns, which is applied in material separation, analysis materials, measuring devices, etc., can solve the problems of protein template molecular embedding, poor selectivity, and inability to recognize and separate proteins, and achieve simple preparation process, easy elution, The effect of low synthesis cost

Active Publication Date: 2009-06-24
中科榆林能源技术运营有限责任公司
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AI Technical Summary

Problems solved by technology

Its protein template molecules are seriously embedded, and the selectivity is poor, so it is impossible to identify and separate proteins
Therefore, neither is suitable for preparing monolithic columns for western blotting

Method used

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  • Production method for protein molecule imprinting integral column
  • Production method for protein molecule imprinting integral column
  • Production method for protein molecule imprinting integral column

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Experimental program
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Embodiment

[0016] Select acrylamide and / or methacrylamide with a weight concentration of 3.0-6.5% and methacrylic acid with a weight concentration of 0.1-0.4% as functional monomers, and a cross-linking agent N, N`-sub Methyl bisacrylamide or piperazine bisacrylamide were co-dissolved in 10 mM phosphate buffer solution (pH 4.5-7.5). Nitrogen gas is passed through to remove dissolved oxygen therein, and then template protein molecule lysozyme, cytochrome C or bovine serum albumin with a weight concentration of 0.15-0.55% is added. Place it at 0-4°C for 2-5 minutes, add initiator ammonium persulfate with a weight concentration of 0.08-0.2% and an accelerator N, N, N`, N`-tetramethylethane with a weight concentration of 0.1-0.15% diamine. After mixing evenly, quickly inject it into the liquid chromatography column, and place it at -5~-20°C for 1~12 hours.

[0017] The weight concentrations of the above substances are their final concentrations in the buffer solution.

[0018] For the pre...

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Abstract

The invention relates to a method for preparing a protein molecular engram block polymer. Functional monomers, crosslinking agents and model protein molecules in proper proportions stand for 2 to 5 minutes at the temperature ranging from 0 to 4 DEG C, and a certain amount of evocating agents and rate accelerating materials are orderly added later. The mixture is put under the environment whose temperature ranges from minus 5 DEG C to minus 20 DEG C after being rapidly injected into a liquid chromatogram column jacket and polyreaction is completed within 1 to 12 hours. The model protein molecules are eluted by dodecyl sodium sulfate whose weight concentration is 10 percent and acetic acid solution whose volume concentration is 10 percent, namely, a macroporous protein molecular engram monolithic column can be prepared. The invention has the advantages as follows: the preparation method is simple and reliable, and the obtained protein molecular engram monolithic column has stronger template molecule recognition property. The technique provides a novel preparation method for developing protein molecular engram monolithic column.

Description

technical field [0001] The invention relates to the preparation of a liquid chromatography column, in particular to a preparation method of a novel protein molecularly imprinted monolithic column. Background technique [0002] Molecular imprinting (molecular imprinting) is a polymer synthesis technology developed in recent years based on the molecular recognition mechanism, which is selective for specific molecules (template molecules or imprinted molecules). Often described as a technique for creating artificial "locks" that recognize "molecular keys". For example, in non-covalent molecular imprinting, the target molecule is usually used as a template, and the molecule is assembled with a polymerizable functional monomer through hydrogen bonding, electrostatic interaction, hydrophobic interaction, etc., and a polymerization reaction occurs in a crosslinking agent. After the reaction, the imprinted molecules are removed to form holes in the polymer with precisely arranged f...

Claims

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Application Information

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IPC IPC(8): G01N30/56
Inventor 张丽华邓启良刘晋湘乔晓强梁振赵鹏张玉奎
Owner 中科榆林能源技术运营有限责任公司
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