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A method of immobilizing protein

A compound and selected technology, applied in the field of immobilizing proteins, can solve the problems of time-consuming, reducing protein activity, denaturation or precipitation of unstable proteins, etc.

Inactive Publication Date: 2018-06-12
PEKING UNIV SHENZHEN GRADUATE SCHOOL
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Despite achieving covalent and directional binding, copper catalysts can cause denaturation or precipitation of some labile proteins, thus potentially reducing protein activity after immobilization
In addition, the introduction of alkyne groups into GST proteins is time-consuming and costly compared to the traditional GSH-GST method

Method used

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  • A method of immobilizing protein
  • A method of immobilizing protein
  • A method of immobilizing protein

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Experimental program
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Embodiment

[0111] In the following examples, all reagents were commercially available and used without further purification. Anhydrous DMF was treated with calcium hydride. Brine refers to a saturated solution of sodium chloride in distilled water.

[0112] The reaction was monitored by TLC performed on 0.25 mm Jiangyou silica gel plates (HSGF254) using UV light as a visualization reagent. Flash column chromatography was performed using Puke silica (ZCX-II).

[0113] 1 H-NMR and 13 C-NMR spectrum is recorded on Bruker Advance400 ( 1 H: 400MHz, 13 C: 100MHz) where the chemical shift values ​​(in ppm) are relative to the following internal standards: TMS (δH0.00 and δC0.00), residual chloroform (δH7.26 and δC77.16), dimethyl sulfoxide (δH2. 50 and δC 39.52), acetone (δH 2.05 and δC 29.92, 206.68), or methanol (δH 3.31 and δC 49.00). 19 F-NMR spectra were recorded on a Bruker Advance400 with CFCl 3 (δF0.00) was used as internal standard. HR-MS was acquired using a Bruker Apex IV RT...

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Abstract

The invention relates to a method for immobilizing proteins and also relates to a compound for immobilizing proteins. The compound has a fluorosulfonyl group, which can be specifically and covalently connected to a tyrosine residue to label the protein.

Description

technical field [0001] The present invention relates to a compound for immobilizing proteins and a method for immobilizing proteins. Background technique [0002] Immobilization of proteins to solid supports such as microarrays, chips, and beads is a powerful method for elucidating protein-protein or protein-ligand interactions and for probing novel protein functions in a high-throughput manner. Unlike the case of DNA microarrays, implementing a general method for protein immobilization presents challenges due to inherent difficulties in achieving selective and uniformly oriented distribution of proteins into solid surfaces. [0003] Conventional immobilization methods mainly rely on three mechanisms: physical attachment, covalent bonding, and bioaffinity immobilization. However, non-selective physical attachment or covalent bonding may block the active site or even denature the protein. Immobilization by co-expression of recombinant fusion tags and coupled proteins can ac...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07D487/04C07D519/00C07K17/02C07K17/04
Inventor 潘峥婴郭天林周怡青
Owner PEKING UNIV SHENZHEN GRADUATE SCHOOL