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Agarose compatible medium for purifying immune globulin of hand-foot-and-mouth disease and preparation method thereof

An immunoglobulin and hand, foot and mouth disease vaccine technology, applied in the biological field, can solve the problems of inability to treat hand, foot and mouth disease, low titer, limited separation accuracy, etc., and achieve excellent biocompatibility, high mechanical strength, high polymer The effect of exclusion limit

Active Publication Date: 2012-11-21
INST OF PROCESS ENG CHINESE ACAD OF SCI +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the separation accuracy of the low-temperature ethanol precipitation method is limited, and only a mixture of various immunoglobulins can be obtained. The product has a low potency and often cannot meet the requirements for the treatment of hand, foot and mouth disease.

Method used

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  • Agarose compatible medium for purifying immune globulin of hand-foot-and-mouth disease and preparation method thereof
  • Agarose compatible medium for purifying immune globulin of hand-foot-and-mouth disease and preparation method thereof
  • Agarose compatible medium for purifying immune globulin of hand-foot-and-mouth disease and preparation method thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0043] Take 100.0 g of washed and drained agarose 6FF gel (GE healthcare company), pour it into a 1L three-necked flask, add 200 ml of 1,4-butanediol-diglycidyl ether, and 100 ml of 0.5M sodium hydroxide solution and 1.5g of sodium borohydride, stirred and reacted at 25°C for 10 hours, after the reaction was over, washed with water, ethanol, and water successively and vacuumed off the solvent to obtain epoxy-activated agarose gel; take 100g of epoxy-activated agar After the sugar gel was washed with water, drained, put into a 1L three-necked flask, added 100ml pH 8.0 hand, foot and mouth disease vaccine (Hualan Biological Co., Ltd.) solution, oscillated in a water bath at 37°C for 24 hours; 1mol / L ethanolamine was shaken and reacted at 37°C for 4 hours; the gel was sequentially washed with deionized water, 0.1M pH 4.0 acetic acid-sodium acetate buffer solution containing 0.5M NaCl, deionized water and 0.1M pH 8.0 boric acid containing 0.5M NaCl - Washing with sodium tetraborat...

Embodiment 2

[0045] Take 100.0 g of washed and drained agarose 6FF gel (GE healthcare company), pour it into a 1L three-necked flask, add 200 ml of 1,4-butanediol-diglycidyl ether, 100 ml of 4M sodium hydroxide solution and 1.5g of sodium borohydride, stirred and reacted at 60°C for 4 hours, after the reaction was completed, washed with water, ethanol, and water in turn and vacuumed off the solvent to obtain epoxy-activated agarose gel; take 100g of epoxy-activated agarose After washing the gel with water, drain it, put it into a 1L three-necked flask, add 200ml pH 14.0 hand, foot and mouth disease vaccine solution (Hualan Biological Co., Ltd.), shake it in a water bath at 37°C for 24 hours; filter it with suction, wash it with deionized water, and add 1mol / L ethanolamine at 37°C for 4 hours; the gel was sequentially washed with deionized water, 0.1M pH4.0 acetic acid-sodium acetate buffer containing 0.5M NaCl, deionized water and 0.1M pH8.0 containing 0.5M NaCl Washing with boric acid-so...

Embodiment 3

[0047] Take 100.0 g of washed and drained agarose 6FF gel (GE healthcare company), pour it into a 1L three-necked flask, add 200 ml of 1,4-butanediol-diglycidyl ether, and 100 ml of 0.5M sodium hydroxide solution and 1.5g of sodium borohydride, stirred and reacted at 60°C for 4 hours, after the reaction was over, washed with water, ethanol, and water successively and vacuumed off the solvent to obtain epoxy-activated agarose gel; take 100g of epoxy-activated agar After the sugar gel was washed with water, drained, put into a 1L three-necked flask, added 500ml pH 8.0 hand, foot and mouth disease vaccine solution (Hualan Biological Co., Ltd.), shaken in a water bath at 37°C for 24 hours; suction filtered, washed with deionized water, added 1mol / L ethanolamine was shaken and reacted at 37°C for 4 hours; the gel was sequentially washed with deionized water, 0.1M pH4.0 acetic acid-sodium acetate buffer containing 0.5M NaCl, deionized water and 0.1M pH8.0 containing 0.5M NaCl boric ...

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Abstract

The invention provides an agarose compatible medium for purifying immune globulin of the hand-foot-and-mouth disease and a preparation method thereof. In the agarose compatible medium, agarose gel is used as a matrix, is activated by an epoxy activating agent, and is coupled with vaccine of the hand-foot-and-mouth disease which is used as petunidin, wherein the agarose gel is made of one of agarose 6FF, agarose 4FF, agarose CL-6B, agarose CL-4B, agarose 6B and agarose 4B. The agarose compatible medium can be used for purifying the immune globulin of the hand-foot-and-mouth disease, has the advantages of good medium selectivity, strong specificity, strong mechanical strength and industrial amplification benefit. Furthermore, the preparation process is simple and is easy to amplify. The medium prepared by the method is adopted for purifying the immune globulin of the hand-foot-and-mouth disease, and has simple and feasible purification method and good separation effect.

Description

technical field [0001] The invention belongs to the field of biotechnology, in particular, the invention relates to an agarose affinity medium used for purifying hand-foot-mouth disease immunoglobulin and its preparation method and application. Background technique [0002] Hand, foot and mouth disease is caused by enteroviruses such as EV71. At the beginning of the onset, there are many small and hard rashes and ulcerative oral inflammation on the hands and feet. The disease mainly occurs in infants and young children. Most patients have mild symptoms, with fever and rashes or herpes on the hands, feet, mouth and other parts as the main features. A small number of patients may be complicated by aseptic meningitis, encephalitis, acute flaccid paralysis, respiratory tract infection and myocarditis, etc., and individual severe cases progress rapidly and are prone to death. Human hand, foot and mouth disease immunoglobulin is a specific immunoglobulin extracted by low-tempera...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C07K16/10C07K1/22
Inventor 苏志国罗坚孔英俊安康范蓓马小伟
Owner INST OF PROCESS ENG CHINESE ACAD OF SCI
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