Affine peptide and application thereof

An affinity peptide and affinity technology, applied in the field of purification and antibody separation, can solve the problem of unsatisfactory purification effect, and achieve the effect of cheap and easy quality control, easy quality control and mature technology.

Active Publication Date: 2019-05-21
NANYANG NORMAL UNIV
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  • Abstract
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  • Claims
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Problems solved by technology

However, when the above-mentioned affinity peptide is used to prepare an affinity medium (the same material as the commercial Protein A / G medium), and when the above-mentioned affinity medium is used for the purification of immunoglobulins by the AKTA protein purification system, the purity of the obtained product is only 90%. %, lower than Protein A / G, the purification effect is not ideal

Method used

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  • Affine peptide and application thereof

Examples

Experimental program
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Effect test

Embodiment 3

[0034] Example 3 Using the affinity medium obtained in Example 2 for the separation and purification of antibodies

[0035] 1) Prepare 10mmol / L Hepes buffer solution, pH7.0, NaCl content 0.15M. Set the flow rate to 1mL / min.

[0036] 2) Use the liquid prepared in 1) as the mobile phase, and run the AKTA protein purification system.

[0037] 3) Take 1 mL of the affinity medium prepared in Example 2 to fill the separation column, and use 1 mL of the Protein A / G prepacked column as a control.

[0038] 4) Load 1mL of antibody fermentation broth respectively. The antibody fermentation liquid can be the antibody fermentation liquid of hybridoma cells or the antibody fusion protein fermentation liquid of Escherichia coli.

[0039] 5) Prepare 0.1M HCl-Gly buffer at pH 2 as the eluent.

[0040] 6) Load 3 mL of the eluate, and collect the eluate.

[0041] 7) Determine the antibody content and purity in the eluent of the antibody fermentation broth, antibody-adsorbed peptide medium, ...

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Abstract

The invention relates to the technical field of antibody separation and purification, in particular to an affine peptide and application thereof. The amino acid sequence of the affine peptide is shownas follows: Glu Top Top Tyr Cys; the affine can be combined with immune globulin for removing or separating or purifying the immune globulin, the effect is equivalent to the effect of Protein A / G, and the safety is better.

Description

technical field [0001] The invention relates to the technical field of antibody separation and purification, in particular to an affinity peptide and its application. Background technique [0002] Antibody is a kind of immunoglobulin secreted by plasma cells that can specifically bind to antigen. Because of its specific binding ability, it can be used as a drug for treating diseases, a reagent for disease diagnosis, and a medium for affinity separation, etc. It has a wide range of applications in the fields of biology, medicine, and chemical industry, and has great social and economic benefits. [0003] The separation and purification of antibodies is the basis of antibody application. At present, the separation and purification of antibodies in industry uses Protein A / G affinity purification medium, which has problems such as expensive medium, easy inactivation, difficult cleaning, and protein A / G shedding pollution. Other separation methods and media are also unsatisfacto...

Claims

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Application Information

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IPC IPC(8): C07K7/06C07K1/22C07K16/00
Inventor 韦宇平柳成宾李娜
Owner NANYANG NORMAL UNIV
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