Method of using nano zeolite molecular sieve assemble material as affinity chromatography filler to separate functional protein molecules

A technology of nano-zeolite and chromatographic filler, which is applied in the field of nano-zeolite molecular sieve assembly material as an affinity chromatographic filler to separate functional protein molecules, can solve the problems of large diffusion resistance, limited use range, narrow pore channels, etc., and achieves uniform pore channels and convenient repeated regeneration. , the effect of stable nature

Inactive Publication Date: 2003-07-23
FUDAN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, for macromolecular systems, due to the narrow pores and large diffusion resistance of traditional zeolite materials, the scope of application is limited.
There are no reports of this type of research method

Method used

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  • Method of using nano zeolite molecular sieve assemble material as affinity chromatography filler to separate functional protein molecules
  • Method of using nano zeolite molecular sieve assemble material as affinity chromatography filler to separate functional protein molecules
  • Method of using nano zeolite molecular sieve assemble material as affinity chromatography filler to separate functional protein molecules

Examples

Experimental program
Comparison scheme
Effect test

example 1

[0040] 0.1 g of self-made modified nano-beta zeolite filler was adjusted to a slurry with an appropriate amount of deionized water, and transferred to a separation column with a bed height of about 1 cm. Then add 1.0ml equilibration buffer solution to equilibrate the column. Dilute 0.50ml to a concentration of 124μgL -1 1.5mgL of histidine or 1.0mL -1 Peptide solution samples (samples are artificially synthesized polypeptide molecules containing different histidine structures) were transferred to the extraction column, and after the liquid flowed out, 1.0ml equilibration buffer solution was added for washing, and the histidine or polypeptide adsorbed on the column passed through Add formic acid-ammonium formate elution buffer solution with pH of 7.0, 6.0, 5.0, and 4.0 in sequence for gradient elution, the elution rate is 10ml / hr, and then detect the absorbance value in each collection tube by ultraviolet spectroscopy, and draw the outflow distribution map. Example 2-5

example 2-5

[0041] Adjust the bed height of the column to be 1.5, 2.0, 2.5, and 3.0 cm respectively, and the other conditions are the same as above, and repeat the above separation experiment. Example 6-8

example 6-8

[0042] Select respectively formic acid-ammonium formate buffer solution system or sodium phosphate-phosphate buffer solution or imidazole gradient solution as the elution system to separate the polypeptide molecule mixture, repeat the example 1 step (the artificially synthesized polypeptide molecules A and B respectively contain different groups amino acid structure, and mixed in a 1:1 molar ratio). The separation of selenoprotein-P in example 9 mouse plasma

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Abstract

In the invention, with being fixed in macropore-micropore multistage ordered zeolite structure through the ion exchnage process, the transition metal ions are as the affiliation active spots where the distinctive combination is occurred with the functional biomacromolecules possesisng the sequential histidine structure. Then, using the gradient elution through the buffer solution to reach the goal of enrich and separation. The carrier provides the advantages of even pore passages, good mechanical and chemical stability, easy to modify the surfaces etc. The diatomaceous earth is as the moldingboard and the molecular sieve of nano Beta zeolite wit surface being modified is as affinity chromatography filler. With the column being filled with the said materials, the satisfying result for separating the functional protein molecules is obtained in different leaching systems.

Description

technical field [0001] The present invention utilizes the nano-zeolite material with macroporous-microporous multi-level ordered structure, and after modifying its surface through methods such as cation exchange, chemical deposition and surface chemical grafting, it is selectively used as a new type of affinity chromatography Separation of functional protein molecules in complex biological systems (such as immobilization of metal ions Co by ion exchange 2+ Afterwards, affinity separation of selenoprotein-P rich in histidine structure in mouse plasma, etc.). Background technique [0002] With the completion of the complete sequencing of the human genome, elucidating the structure and function of its encoded proteins has become a new frontier in life sciences. Around 1995, some far-sighted scientists timely proposed the concept of "post-genome". That is, after the static base sequence of the genome is clarified, the study of the dynamic biological function of the genome is t...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): B01D15/08C07K1/22G01N30/62G01N30/74
Inventor 徐芳王亚军王德举唐颐杨芃原
Owner FUDAN UNIV
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