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Bacterial strain for preparing high efficiency capsule glycoprotein immunomodulator, screening and application thereof

An immunomodulator, capsular saccharide technology, applied in peptide/protein components, antibacterial immunoglobulins, biochemical equipment and methods, etc. Denaturation inactivation and high immune activity

Inactive Publication Date: 2007-06-06
TIANJIN MEDICAL UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0012] In view of the above situation, combined with modern biotechnology, the present invention overcomes the deficiencies in the prior art and solves the problem that the current Klebsiella pneumoniae capsular glycoprotein production strains do not have the characteristics of the Chinese Klebsiella pneumoniae infection epidemic strains, To make the immunomodulatory substances more suitable for the domestic infected population, provide a strain of Klebsiella pneumoniae Kp-9 that produces high-efficiency capsular glycoprotein, as well as a screening method for Klebsiella pneumoniae strains and a new preparation method for capsular glycoprotein

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0054] Example 1: A screening method for Klebsiella pneumoniae producing high-efficiency capsular glycoprotein. It includes the following steps:

[0055] (1) Bacterial source selection: domestic clinically isolated Klebsiella pneumoniae strain and Klebsiella pneumoniae standard strain 46114 were selected as bacterial sources for screening and comparison.

[0056] (2) Preparation of Klebsiella pneumoniae capsular glycoprotein antiserum: 5 mg / mL Klebsiella pneumoniae capsular glycoprotein was used as immunogen, and a dose of 0.3 mL / body was used to immunize male rats without basic cross-antibody. Eared white rabbits, blood was collected from the immune white rabbits, and the immune serum was obtained by centrifuging at 4000rpm for 20 minutes.

[0057] (3) Screening scheme: suspend the bacterial culture in the ELISA coating solution (turbidity reaches 10 7 CFU / mL), this bacterial suspension was coated on an ELISA plate, 100 μL per well, placed at 35°C for 3 hours, and then plac...

Embodiment 2

[0058] Example 2: A screening method for Klebsiella pneumoniae producing high-efficiency capsular glycoprotein. It includes the following steps:

[0059] (1) Bacterial source selection: select domestic clinically isolated Klebsiella pneumoniae strain and Klebsiella pneumoniae standard strain 46114 as the bacterial source for screening and comparison;

[0060] (2) Preparation of Klebsiella pneumoniae capsular glycoprotein antiserum: use 1 mg / mL Klebsiella pneumoniae capsular glycoprotein as the immunogen, and immunize male big ears without basic cross-antibody at a dose of 0.1 mL / body White rabbits, then blood was collected from the immune white rabbits, and centrifuged at 3000rpm for 20 minutes to obtain immune serum;

[0061] (3) Screening scheme: suspend the bacterial culture in the ELISA coating solution (turbidity reaches 10 5 CFU / mL), this bacterial suspension was coated on an ELISA plate, 100 μL per well, placed at 30°C for 2 hours, and then placed at 4°C for 10 hours,...

Embodiment 3

[0062] Example 3: A screening method for Klebsiella pneumoniae producing high-efficiency capsular glycoprotein. It includes the following steps:

[0063] (1) Bacterial source selection: select domestic clinically isolated Klebsiella pneumoniae strain and Klebsiella pneumoniae standard strain 46114 as the bacterial source for screening and comparison;

[0064] (2) Preparation of Klebsiella pneumoniae capsular glycoprotein antiserum: use 10 mg / mL Klebsiella pneumoniae membrane glycoprotein as the immunogen, and immunize male big ears without basic cross-antibody at a dose of 0.5 mL / body White rabbits, then blood was collected from the immune white rabbits, and the immune serum was obtained by centrifugation at 5000rpm for 20 minutes;

[0065] (3) Screening scheme: suspend the bacterial culture in the ELISA coating solution (turbidity reaches 10 9 CFU / mL), this bacterial suspension was coated on an ELISA plate, 100 μL per well, placed at 40°C for 4 hours, and then placed at 4°C...

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Abstract

The invention discloses a bacterial strain for preparing highly effective capsula glucoprotein immuno-modulatory agent, its selection and use thereof, which relates to a strain of K. peneumoniae and its screen selecting method, and the method for preparing capsula glucoprotein by using the strain, wherein the K. peneumoniae capsula glucoprotein immuno-modulatory agent is prepared through the steps of strain screening, culture, cracking, crude extracting and purification, thus overcoming the problem of drug tolerance for antibiotics when used for treating infection.

Description

technical field [0001] The invention belongs to the technical field of strain screening and preparation of bacterial active ingredients, and in particular relates to a high-efficiency capsular glycoprotein-producing Klebsiella pneumoniae and a screening method thereof, and a method for preparing capsular glycoprotein using the strain. Background technique [0002] At present, the study of immunomodulators is a hot topic, and cytokines are a very active research field in immunology in recent years. Many factors such as IL-2, GM-CSF, TNF-α, etc. have been clinically proven to be It has a good regulatory effect, but cytokines have the following problems as immunomodulators: [0003] 1) It needs to be synthesized by gene recombination technology, and the price is relatively expensive; [0004] 2) The half-life is short, and the retention time in the body is short, so the duration of the drug effect is short; [0005] 3) The administration of supraphysiological dosage has great...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20C12P21/00C07K14/26C07K16/12C07K1/36A61K38/00A61K39/395G01N33/53G01N30/00C12R1/22A61P31/04A61P37/02
Inventor 陈锦英
Owner TIANJIN MEDICAL UNIV
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