Unlock instant, AI-driven research and patent intelligence for your innovation.

New alginate monoclone antiboeg and its preparation method and use

A technology of monoclonal antibody and alginate, which is applied in the direction of anti-bacterial immunoglobulin, cells modified by introducing foreign genetic material, fusion cells, etc. It can solve the problems of lack of in-depth research reports and difficulties in the means of biological effects of alginate, etc. question

Inactive Publication Date: 2004-10-20
中国人民解放军军医进修学院
View PDF0 Cites 0 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are difficulties in the detection and determination of alginate, and the means of blocking the biological effect of alginate.
There is no definite treatment for Pseudomonas aeruginosa biofilm-related infections. Foreign scholars have proposed that alginate monoclonal antibodies may have a certain inhibitory effect on bacterial biofilms (Mai GT, McCormack JG, Seow WK, et al.Inhibition of adherence of mucoid Pseudomonas aeruginosa by alginase, specific monoclonal antibodies, and antibiotics. Infect Immun 1993Oct; 61(10): 4338-43), but no in-depth research reports

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] Example 1 Preparation of immunogen: Alginate cross-linked with BSA

[0024] Precisely weigh 5.0 mg of alginate and dissolve in 0.05M 50ml of boric acid buffer at pH 10, add EDCI100mg under a magnetic stirrer and react at room temperature for 8 hours, then add 0.05M 5.0ml of boric acid buffer at pH 10 to dissolve BSA, and react at room temperature for 8 hours Dialyze overnight at 4°C. EDCI should be freshly prepared.

Embodiment 2

[0025] Example 2 Animal Immunization

[0026] The immunogen prepared as in Example 1, that is: the alginate-BSA complex was dissolved in physiological saline, mixed with an equal volume of Freund's complete adjuvant, and BALB / C mice (male, six-week-old, 18-20 g , provided by the Animal Center of Chinese Academy of Medical Sciences) subcutaneously injected into the abdomen at multiple points, 40 μg / 200 μl / animal. The first booster immunization was carried out two weeks after the basic immunization, and then every 2-4 weeks, with a dose of 20μg / 200μl / bird.

[0027] One day after the initial immunization and 6 booster immunizations, the antiserum titer of the fusion mice to be taken from the spleen was measured by indirect ELISA. The normal mouse serum was used as the negative control, and the determination was carried out on the basis of (measurement well A value-blank value) / (negative control well A value-blank value)≥2.1.

[0028] The endpoint titer of the antiserum used for...

Embodiment 3

[0029] Example 3 Cell Fusion

[0030] Cell fusion adopts the polyethylene glycol method: take the spleen of the mouse immunized as described in Example 2 aseptically, make a cell suspension, centrifuge, count the cells, and splenocytes (about 1 × 10 8 ) and SP2 / 0 cells (about 1×10 7 ) were mixed, centrifuged at 1000r / min for 10min, and the supernatant was removed. Place the centrifuge tube in a 37°C water bath, slowly add 1ml of 50% PEG solution (Sigma), let it stand for 1 minute, and then slowly add 9ml of serum-free culture medium IMDM. Centrifuge at 800r / min for 10min, discard the supernatant, wash twice, add complete medium to resuspend the cells for inoculation.

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Affinityaaaaaaaaaa
Login to View More

Abstract

A novel monoclonal antibody of alginate, the hybrid tumor clone secreting said monoclonal antibody, its preparing process and its application are disclosed.

Description

field of invention [0001] The present invention relates to a novel alginate monoclonal antibody and a hybridoma clone secreting the monoclonal antibody, and also relates to a preparation method of the monoclonal antibody and an application of the monoclonal antibody. Background of the invention [0002] Alginate is one of the important pathogenic factors secreted by mucus-type Pseudomonas aeruginosa, which plays an important role in the adhesion of bacteria to biomedical materials or body cavity, the formation of biofilm, and the resistance to body immunity. effect. When Pseudomonas aeruginosa touches the surface of an object, it triggers the biosynthesis of alginate, promotes its adhesion, makes the bacteria aggregate to a certain number, forms colonies, and finally leads to the formation of biofilm (Costerton J.W.Overview of microbial biofilms.J - Ind-Microbiol. 1995 Sep;15(3):137-40). Biofilm-associated infection caused by the formation of biofilm by bacteria is one of ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): C07K16/12C12N5/12C12P21/08G01N33/577
Inventor 王睿裴斐柴栋方翼陈迁方向群
Owner 中国人民解放军军医进修学院