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Process for preparing acellular short corynebacteria preparation

A Corynebacterium brevis, cell-free technology, used in bacteria, bacterial antigen components, drug combinations, etc., can solve problems such as cell fragmentation, achieve uniform particle size, improve absorption and diffusion, and reduce fever.

Inactive Publication Date: 2004-01-07
上海昌润生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although there are many methods for cell disruption, it is difficult to disrupt cells into nanoscale with uniform particle size.

Method used

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  • Process for preparing acellular short corynebacteria preparation
  • Process for preparing acellular short corynebacteria preparation

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0048] According to the method described in the 2000 edition of "China Biological Products Regulations", the establishment, verification and other verification of bacterial seed batches were carried out.

[0049] The medium used in this example is the thioglycolate medium (without agar) produced by Beijing Sanyao Science and Technology Development Company, and its ingredients are: tryptone 15g / l, yeast extract powder 5g / l, glucose 5g / l , sodium chloride 2.5g / l, L-cystine 0.5g / l, sodium thioglycolate 0.5g / l. The medium was filtered through a 0.22 μm membrane before use.

[0050] Open the CP (Propionibacterium acnes, scientific name, propionibacterium acnes) 771 (see "China Biological Products Regulations") strain tube, each tube contains 6 billion bacteria, and draw about 0.2-0.3ml of thioglycolate medium (not containing agar), add it to the bottom of the strain tube to dissolve it completely, suck it out, add it to the middle tube (capacity 38ml), mix it with 8-10ml of the ab...

Embodiment 2

[0055] Referring to the method of Example 1, CP (scientific name propionibacterium acnes, propionibacterium acnes) 65101 (76-27) was inoculated in a nutrient tank after 4 consecutive passages, and was cultured in a thioglycolate medium (without agar) (before use) Filter the culture medium with a 0.45 μm membrane) and culture at 36-37°C for 6 days; check the bottles one by one under the microscope, and discard those contaminated by bacteria; combine and collect the bacteria without bacteria contamination, heat and boil for 60 minutes, and use ultra-high pressure jet Collider at 1500kgf / cm 2 Broken bacterium under pressure 5 times; The broken suspension was centrifuged at 8000rpm for 1 hour, washed with sterile normal saline for 5 times, sterility test (2000 edition "China Biological Products Regulation") and pyrogen detection (2000 edition Pharmacopoeia of the People's Republic of China ") is qualified to make a suspension with a solid content of 2.0mg / ml, and heat at 65°C for ...

Embodiment 3

[0059] With reference to the method of Example 1, CP (Propionibacterium acnes) 65102 (H-84) was inoculated in a nutrient tank after 4 consecutive passages, and cultured in a peptone medium at 36-37°C for 7 days; bottle by bottle Microscopic examination, those with contamination by miscellaneous bacteria were discarded; combined and collected bacterial cells without contamination by miscellaneous bacteria were heated and boiled for 30 minutes. 2 Broken thalli under pressure 10 times; The suspension after broken is centrifuged at 12000rpm, washes precipitate 4 times with sterile normal saline, sterility test (2000 version "Chinese Biological Products Regulations") and pyrogen detection (2000 version "People's Republic of China Regulations") Pharmacopoeia ") qualified to make a suspension with a solid content of 1.0mg / ml, heated at 65°C for 100 minutes to obtain NCP.

[0060] Detect (solid content 1mg / ml) NCP pyrogen 60EU / ml by limulus reagent method (2000 edition "Pharmacopoeia ...

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Abstract

The present method of non-cell corynebacterium parvum (NCP) preparation includes the following steps: continuous 2-4 times of passage of corynebacterium parvum (CP) working seed batch strain, then inoculating in large flask or nutrient tank, culturing for 5-7 days in proper culture medium, collecting thallus having no heterobacteria pollution, heating, boiling and inactivating to obtain bacetrialsolution, breaking bacterial solution, centrifugation and collecting precipitate, washing to obtain suspension, heating said suspension and sterilizing so as to obtain the invented NCP. Said invention also provides its application function.

Description

technical field [0001] The invention relates to a preparation method of a biological product, in particular to a preparation method of an acellular short coryneform bacterium preparation (NCP), belonging to the field of biological products. Background technique [0002] Corynebacterium breve bacterin (CPV, 1995 edition of "China Biological Products Regulations") or short Corynebacterium preparations (CPP, 2000 edition of "China Biological Products Regulations") is a non-specific immunomodulator. It is a dead bacterin vaccine made from Corynebacterium brevis (CP) inactivated by formaldehyde, and its preparation process is disclosed in the 1995 and 2000 editions of "China Biological Products Regulations". Currently used CPP preparations also include inactivated vaccines produced by Merleux Research Institute in France; inactivated vaccines produced by Wellcome Pharmaceuticals in the UK, etc. [0003] CPP has the ability to activate the mononuclear phagocyte system. The role ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/05A61P35/00C12N1/20
Inventor 高尚先李守悌
Owner 上海昌润生物科技有限公司
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