A process for extracting lipopolysaccharide from Salmonella paratyphi A by cold phenol method

A Salmonella lipid and paratyphoid technology, which is applied in the field of pharmaceutical preparation, can solve the problems of high residual protein content, influence hydrolysis yield, insufficient LPS purity, etc., and achieve the effects of improving purity, improving purity and safety.

Active Publication Date: 2020-06-09
罗益(无锡)生物制药有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the LPS extracted by this extraction method often has a high residual protein content, and the purity of LPS is not enough, which will affect the subsequent hydrolysis yield.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Embodiment 1: a kind of technique utilizing cold phenol method to extract the lipopolysaccharide of Salmonella paratyphi A, described extraction technique comprises the steps:

[0024] Step a: Bacteria treatment: After weighing an appropriate amount of bacteria, add water for injection in proportion, and smash the bacteria;

[0025] Step b: the first extraction: adding phenol to the cells treated in step a, heating in a water bath for 1-3 h, then placing the obtained solution in a cold storage overnight, and then centrifuging to collect the supernatant;

[0026] Step c: the second extraction: add cold phenol to the supernatant obtained in step b, shake for 8-20 minutes, centrifuge, collect the supernatant, and place the supernatant in a dialysis bag for dialysis;

[0027] Step d: Extraction of LPS: Add MgCl to the dialyzed solution in step c 2 and NaAc to a final concentration of 0.1-0.3 mol / l MgCl 2 and 0.02-0.05mol / l NaAc; then add ethanol to it to a final concentra...

Embodiment 2

[0028] Embodiment 2: a kind of technique utilizing cold phenol method to extract lipopolysaccharide of Salmonella paratyphi A, described extraction technique comprises the steps:

[0029] Step a: Bacteria treatment: After weighing an appropriate amount of bacteria, add the injection water according to the dosage ratio of bacteria and injection water: (8-10), and crush the bacteria;

[0030] Step b: the first extraction: add 90% phenol to the bacteria treated in step a, the ratio of the amount of phenol to the amount of bacteria is (1-2): (1-3); and at 60 Heating in a water bath at -70°C for 1-3 h, then placing the obtained solution in a cold storage overnight, and then centrifuging to collect the supernatant;

[0031] Step c: the second extraction: add cold phenol at a concentration of 70% to the supernatant obtained in step b, and the dosage ratio of cold phenol to the supernatant is (1-2): (1-3); shake Centrifuge after 8-20 minutes, collect the supernatant, and place the su...

Embodiment 3

[0033] Embodiment 3: utilize cold phenol method to extract the technology of Salmonella paratyphi A lipopolysaccharide, this technology specifically comprises the following steps:

[0034] Step a: Bacteria treatment: After weighing an appropriate amount of bacteria, add water for injection at a ratio of 1:10, and crush the bacteria;

[0035] Step b: the first extraction: add 90% phenol to the cells treated in step a at a ratio of 1:1, heat in a water bath at 68°C for 1 hour, then place the obtained solution in a refrigerator overnight, and then centrifuge Separate and collect the supernatant;

[0036] Step c: the second extraction: add 70% cold phenol to the supernatant obtained in step b at a ratio of 1:1, shake for 10 minutes and then centrifuge to collect the supernatant, and place the supernatant in dialysis in-bag dialysis;

[0037] Step d: Extraction of LPS: Add MgCl to the dialyzed solution in step c 2 and NaAc to a final concentration of 0.1 mol / l MgCl 2 and 0.03mo...

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Abstract

The invention discloses a process for extracting salmonella paratyphi A lipopolysaccharide by utilizing a cold phenol method. The extracting process comprises the following steps that (a) thallus treatment is performed; (b) first-time extraction is performed; (c) second-time extraction is performed; (d) LPS extraction is performed. In the mode, LPS purity can be improved. Compared with the prior art, the LPS purity is improved on the premise that new devices and reagents are not increased and the final product yield is not influenced, and the purity, safety and effectiveness of a final product are improved.

Description

technical field [0001] The invention relates to the field of medicine preparation, in particular to a process for extracting lipopolysaccharide from Salmonella paratyphi A. Background technique [0002] With the development of medical technology and the wide application of typhoid Vi polysaccharide vaccine, the incidence of typhoid fever has gradually decreased, but the epidemic of type A paratyphoid (paratyphoid) has emerged, and enteric fever caused by paratyphoid fever has been obvious both at home and abroad in recent years. The rising trend of the disease and the continuous increase in the number of patients have attracted more and more attention. [0003] Paratyphoid A is an intestinal infectious disease, which can be spread through domestic water, food, contact with flies, cockroaches, etc. to cause epidemics. Symptoms: general malaise at the beginning, poor appetite, headache; fever, body temperature up to 40°C within 5 to 6 days, persistent; dull expression, red ra...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C08B37/00
CPCC08B37/0003
Inventor 周富昌
Owner 罗益(无锡)生物制药有限公司
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