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Biomimetic affinity purification method of vitellus immune globulin

A technology of egg yolk immunoglobulin and purification method, applied in the direction of egg immunoglobulin and the like, can solve the problems of high cost, cumbersome steps, unstable polypeptide properties, etc., and achieves reduction of large-scale purification steps, reduction of production costs, The effect of improving purification efficiency

Inactive Publication Date: 2006-03-29
上海荣君生物医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The preparation of TG19318 requires solid-phase peptide synthesis and high-performance liquid chromatography purification, and then immobilized on the medium, the steps are cumbersome and the cost is high; TG19318 itself is an unstable peptide and cannot tolerate online cleaning conditions
Although the use of TG19318 can simplify the complex IgY purification process, the above shortcomings limit the use of TG19318 for large-scale purification applications

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] (1) Preparation of separation material.

[0018] Take NH 2 -Sepharose (200ml), washed with 5 times the volume of 1M NaCl, then fully washed with distilled water, drained of water, poured into a reaction vessel, added 150ml of water, and placed in an ice-salt bath for precooling. When the temperature dropped to 5°C, start stirring. Dissolve triazoxide (40 g) with pre-cooled acetone and add to the reaction vessel. with saturated NaHCO 3 Keep the pH of the solution between 6 and 7, react at 5°C for 3 hours, take out, 3×10 times the volume of water / acetone (1:1, 1:3, 0:1, 1:1, 3:1, 1:0) and washed successively to obtain 1-amino-Sepharose-3,5-dichloro-2,4,6-triazoxide (190ml).

[0019] Get 1-amino-Sepharose-3,5-dichloro-2,4,6-triazoxide (180ml), weigh aminobenzoic acid (20g) and dissolve it with deionized water (50ml), and dissolve it with 1-amino-Sepharose - Mix 3,5-dichloro-2,4,6-triazazine B, place at a temperature of 50° C. and stir for 24 hours. After the reaction...

Embodiment 2

[0025] (1) Preparation of separation material.

[0026] Take NH 2 -Sepharose (200ml), washed with 5 times the volume of 1M NaCl, then fully washed with distilled water, drained of water, poured into a reaction vessel, added 150ml of water, and placed in an ice-salt bath for precooling. When the temperature dropped to 5°C, start stirring. Dissolve triazoxide (40 g) with pre-cooled acetone and add to the reaction vessel. with saturated NaHCO 3Keep the pH of the solution between 6 and 7, react at 5°C for 3 hours, take out, 3×10 times the volume of water / acetone (1:1, 1:3, 0:1, 1:1, 3:1, 1:0) and washed successively to obtain 1-amino-Sepharose-3,5-dichloro-2,4,6-triazoxide (190ml).

[0027] Get 1-amino-Sepharose-3,5-dichloro-2,4,6-triazoxide (180ml), weigh aminobenzoic acid (20g) and dissolve it with deionized water (50ml), and dissolve it with 1-amino-Sepharose - Mix 3,5-dichloro-2,4,6-triazazine B, place at a temperature of 50° C. and stir for 24 hours. After the reaction ...

Embodiment 3

[0033] (1) Preparation of separation material.

[0034] Take NH 2 -Sepharose (200ml), washed with 5 times the volume of 1M NaCl, then fully washed with distilled water, drained of water, poured into a reaction vessel, added 150ml of water, and placed in an ice-salt bath for precooling. When the temperature dropped to 5°C, start stirring. Dissolve triazoxide (40 g) with pre-cooled acetone and add to the reaction vessel. with saturated NaHCO 3 Keep the pH of the solution between 6 and 7, react at 5°C for 3 hours, take out, 3×10 times the volume of water / acetone (1:1, 1:3, 0:1, 1:1, 3:1, 1:0) and washed successively to obtain 1-amino-Sepharose-3,5-dichloro-2,4,6-triazoxide (190ml).

[0035] Get 1-amino-Sepharose-3,5-dichloro-2,4,6-triazoxide (180ml), weigh aminobenzoic acid (20g) and dissolve it with deionized water (50ml), and dissolve it with 1-amino-Sepharose - Mix 3,5-dichloro-2,4,6-triazazine B, place at a temperature of 50° C. and stir for 24 hours. After the reaction...

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Abstract

A bionic affinity purifying process for vitelline immunoglobulin includes such steps as activating the basic chromatographic medium by reacting on trichlorotrinitroazine, respectively reacting on p-amino benzoic acid, tyrosine and arginine to obtain bionic affinity separating material, preparing affinity chromatographic column, chromatography of vitelline immunoglobulin specionent, flushing, and eluting. Its advantages are high purity and high recovery rate.

Description

technical field [0001] The invention relates to a method in the field of biotechnology, in particular to a method for biomimetic affinity purification of yolk immunoglobulin. Background technique [0002] IgY (yolk antibody or yolk immunoglobulin) is also found in the yolk of birds, reptiles, amphibians, and fish, and a short, sheared IgY(ΔFc) (Immunol Today, 1995, 16(8), 392-8.). In terms of immunodiagnosis, since IgY does not activate complement, bind rheumatoid factor, bind protein A, protein G, mammalian Fc receptors and complement, and has little cross-reaction with IgG, it can be used as an immunological laboratory detection tool. Used to determine circulating complexes, rheumatoid factors and complement, etc., to reduce false positives (J Immunol Met, 1992, 156, 79-83.) Yolk immunoglobulin plays an important role in the field of analysis, diagnosis, disease prevention and treatment of diseases. Compared with mammalian antibodies, egg yolk immunoglobulin has many obv...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/02
Inventor 李荣秀刘浩然
Owner 上海荣君生物医药科技有限公司
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