Small molecular antibody affinity peptide and application thereof

A small molecule antibody and affinity peptide technology, applied in the field of molecular biology, can solve the problems of protein A instability, easy shedding of ligands, poor selectivity, etc.

Inactive Publication Date: 2014-11-26
INST OF PROCESS ENG CHINESE ACAD OF SCI
View PDF2 Cites 10 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] 1) Expensive: Because the protein A ligand needs to be produced and purified by genetic engineering, the production cost is high, and its price reaches $10,000/liter of affinity medium;
[0005] 2) There is a lack of effective cleaning methods. Since protein A is unstable in an alkaline environment, general sodium hydroxide in-situ cleaning methods cannot be used;
[0006] 3) Since the ligand is also a protein, it is easily hydrolyzed by the protease present in the raw material solution, thereby reducing the separation effect;
[0008] 5) The ligand is easy to fall off, and the fallen protein A will cause human immune response and has been clinically proven

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Small molecular antibody affinity peptide and application thereof
  • Small molecular antibody affinity peptide and application thereof
  • Small molecular antibody affinity peptide and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 2I

[0040] Embodiment 2 IgG affinity property determination

[0041] 1) Prepare 10mmol / L Hepes buffer, pH 7.4, NaCl content 150mmol / L.

[0042] 2) Use the liquid prepared in 1) as the mobile phase, and run the BiacoreT200 analyzer.

[0043] 3) Insert the detection chip CM5 and set the flow rate to 30 μL / min.

[0044] 4) Inject 100 μL each of EDC and NHS in sequence to activate the chip.

[0045] 5) Dissolve IgG in sodium acetate buffer, pH 4.5, IgG 100μg / mL.

[0046] 6) Inject 100 μL of the IgG solution in 5), and immobilize IgG on the surface of the chip.

[0047] 7) Small molecule antibody affinity peptide 1 (amino acid sequence shown in SEQ ID No.11), small molecule antibody affinity peptide 2 (amino acid sequence shown in SEQ ID No.34) and natural ligand (ProG and ProA), dissolved in 1) to prepare liquid, content 100μg / mL.

[0048] 8) Sequentially inject different concentrations of peptides and natural ligands (concentrations from high to low are 500 μm, 250 μm, 125 μm, 6...

Embodiment 3

[0053] Example 3 Comparison of Adsorption Properties of Small Molecule Antibody Affinity Peptides with IgG and HAS

[0054] 1) Prepare 10mmol / L Hepes buffer, pH 7.4, NaCl content 150mmol / L.

[0055] 2) Use the liquid prepared in 1) as the mobile phase, and run the BiacoreT200 analyzer.

[0056] 3) Insert the detection chip CM5 and set the flow rate to 30 μL / min.

[0057] 4) Inject 100 μL each of EDC and NHS in sequence to activate the chip.

[0058] 5) Dissolve IgG in sodium acetate buffer, pH 4.5, IgG 100μg / mL. Also dissolve HSA in sodium acetate buffer, pH 4.5, HSA 100 μg / mL.

[0059] 6) Inject 100 μL of the IgG solution in 5), and immobilize IgG on the surface of the chip.

[0060] 7) Inject 100 μL of the HSA solution in 5), and immobilize HSA on the surface of another chip.

[0061] 8) Dissolve small molecule antibody affinity peptide 3 (amino acid sequence shown in SEQ ID No.1) and small molecule antibody affinity peptide 4 (amino acid sequence shown in SEQ ID No.4) ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

No PUM Login to view more

Abstract

The invention relates to the field of molecular biology, in particular to a small molecular antibody affinity peptide with high antibody selectivity and application. The amino acid sequence of the small molecular antibody affinity peptide disclosed by the invention is shown as any of SEQ ID NO.1 to SEQ ID NO.54. The small molecular antibody affinity peptide according to the invention is combined with the Fc segment of immunoglobulin G (IgG), and is not combined with human serum albumin (HAS). When the small molecular antibody affinity peptide disclosed by the invention is taken as an affinity ligand, efficient, safe and economical IgG separation and purification can be realized.

Description

technical field [0001] The invention relates to the field of molecular biology, in particular, the invention relates to an antibody high-selectivity small molecule antibody affinity peptide and its application. Background technique [0002] Immunoglobulin IgG, that is, an antibody is a special protein molecule, which is used as a reagent for in vitro diagnosis, a drug for treating diseases, a ligand for immunoaffinity chromatography, etc., and has a wide range of applications in the fields of life science research, biotechnology and medicine. Applications. [0003] At present, the separation and purification of IgG mostly use the natural affinity ligand Protein A / G of the antibody. Due to the ability of Protein A / G to specifically recognize the Fc fragment of the antibody, the protein A / G affinity chromatography is used to purify the antibody, and the obtained The purity of the product is high. However, since Protein A / G is also an active biological substance, there are al...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
IPC IPC(8): C07K5/103C07K5/113C07K1/22A61M1/34A61M1/18
Inventor 徐霞韦宇平徐建栋
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap