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Method for preparing antibiotic peptide and transfer factor by in vitro culture and induction of lymphocyte

A lymphocyte and transfer factor technology, applied in fermentation and other directions, can solve the problems of less production raw material sources, complex production process, low output rate, etc., and achieve the effects of saving manpower, simple production process and high output rate.

Active Publication Date: 2006-05-03
DALIAN SANYI ANIMAL MEDICINE CO LTD +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0008] The present invention aims to solve the technical problems existing in the prior art such as complicated product production process, low output rate, high cost, less sources of production raw materials, and the potential risk of carrying exogenous viruses and other pathogenic microorganisms in the product, and provides a A method for producing antibacterial peptides and transfer factors by inducing lymphocytes to prepare antimicrobial peptides and transfer factors by in vitro culture that can avoid products carrying exogenous viruses and other pathogenic microorganisms with simple production process, high output rate, low cost, and less raw material consumption, and can be subcultured for multiple times Production

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0019] 1. Using fresh pig spleen as raw material, a monolayer of lymphocytes was prepared according to a conventional method.

[0020] The specific operation method can be carried out as follows:

[0021] a. Processing fresh pig spleen

[0022] Take fresh pig spleen, put it into a sterile beaker immediately, add balanced salt solution to soak for half an hour or immerse in 1% bromogeramine solution, take out the spleen surface with iodine alcohol to disinfect, use dissecting scissors and tweezers to remove spleen fat and capsule, Wash once with balanced salt solution, then cut the spleen into small pieces and transfer them to a plate, and finally cut the spleen into 1mm pieces with ophthalmic scissors 3 Small pieces, then washed 2-3 times with balanced salt solution to remove blood cells, pigment substances and cells damaged mechanically during the shredding process;

[0023] b. Digest and disperse tissue pieces

[0024] Suck off the balanced salt solution washed in the pre...

Embodiment 2

[0047] 1. Using animal peripheral blood as a raw material, a lymphocyte monolayer was prepared according to a conventional method.

[0048] The specific method is: take 20 ml of animal whole blood in a sterile vein, add 0.2 ml of 1% heparin solution for anticoagulation, let it stand for about 60 minutes, and use a straw with a rubber ball to absorb all the plasma above the red blood cell layer, especially immediately above the red blood cell layer. The white blood cell layer should be absorbed as much as possible, divided into sterilized centrifuge tubes, washed repeatedly with balanced salt solution for 2 to 3 times, and the centrifugal speed is 800 to 1200 rpm, and then cultured with 40 ml of hydrolyzed milk protein containing 30% calf serum , mixed evenly and divided into 100ml culture square bottles or brick bottles, tightly stoppered, placed in a 37°C incubator or brick bottle machine for cultivation, after 2 to 3 days, the white blood cells evenly adhered to the wall, and...

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Abstract

The invention discloses a method to prepare antibacterial peptide and transfer factor by culturing and inducing lymphocyte in vitro, which comprises: preparing and subculturing lymphocyte single layer by spleen or peripheral blood; using canavaline A to induce and culture the single layer, and obtaining the product. This invention needs just supersonic wave and frost-thaw treatment without purification, can apply the product directly on animal for antibiosis and antiviral, needs not to slay lots of live pigs, has high yield with low cost, and can prevent the product bringing extraneous virus.

Description

Technical field: [0001] The invention relates to a production method of antibacterial peptides and transfer factors, in particular a method for preparing antibacterial peptides and transfer factors by in vitro culture-induced lymphocytes with simple production process and high output rate. Background technique: [0002] Antibacterial peptides (Anti-bacterial peptides, referred to as ABP) are induced and produced by the animal immune defense system, which can resist exogenous pathogens. It has small molecular weight and strong activity. The protozoan effect, even has a killing effect on cancer cells, and is widely used. The current preparation method uses fresh pig small intestines as raw materials, removes fat, serosa and contents, weighs them, freezes them, cuts them into pieces, and smashes them into pulp. After homogenization, boil in water for 15 minutes, add 5% acetic acid at a ratio of 1:1, and stir overnight at 4°C, and centrifuge the mixture at 8,000 rpm for 30 minu...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12P21/02
Inventor 江国托
Owner DALIAN SANYI ANIMAL MEDICINE CO LTD
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