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Human source anti- tetanus exotoxin antibody and preparation method and use thereof

A technology for tetanus exotoxin and tetanus toxoid, which is applied in the fields of botanical equipment and methods, biochemical equipment and methods, antibodies, etc., and can solve problems such as insufficient blood source

Inactive Publication Date: 2005-03-16
北京明新高科技发展有限公司 +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The human anti-tetanus toxin antibody prepared by this method can avoid allergic reactions caused by heterogeneous serum proteins, and can overcome the problems of insufficient blood source and pathogen contamination in the production of human immunoglobulins

Method used

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  • Human source anti- tetanus exotoxin antibody and preparation method and use thereof
  • Human source anti- tetanus exotoxin antibody and preparation method and use thereof
  • Human source anti- tetanus exotoxin antibody and preparation method and use thereof

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Embodiment Construction

[0109] Phagemid pComb3 (4000bp) was purchased from Scripps Research Institute, USA; XL1-Blue and helper phage VCSM13 were purchased from Stratagene Corporation, USA; Trizol was purchased from Gibco-BRL Corporation; PCR kits, restriction enzymes, and T4 DNA ligase were purchased from In Promega; tetanus exotoxin and standard human tetanus immunoglobulin (TIG) were purchased from China National Institute for the Control of Pharmaceutical and Biological Products. Goat anti-human IgG Fab, inducer IPTG (isopropyl-β-d-thiogalactoside), chromogen OPD (o-phenylenediamine), and 3-morpholine propanesulfonic acid (MOPS) were all from Sigma product. Yeast and CHO were purchased from Promega.

[0110] 1. PCR amplification of human IgG Fab segment gene:

[0111] Lymphocytes were isolated from the anticoagulated blood of volunteers immunized with tetanus toxoid for 6 days with lymphocyte separation medium, total cellular RNA was extracted with Trizol, and the extracted RNA was reverse-tran...

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Abstract

The invention discloses a human source anti-tetanus exotoxin antibody (HTAT-Fab) and preparation method, comprising: constructing human immunity phage antibody bank, screening phage positive clone, further getting HTAT-Fab gene with a specific neutralization activity and high affinity. The gene can be expressed in the procaryotic cell such as E.coli, eukaryotic cell such as microzyme, or mammalian cell such as CHO, purifying to get highly purified HTAT-Fab with a strong tissue penetrability, a high affinity. The HTAT-Fab product can not only eliminate the allergic reaction generated by horse serum anti-tatanus antitoxin (TAT) (foreign protein), but also avoid the blood source for producing human tetanus immunoglobulin (HTIG) and the latent virus pollution.

Description

field of invention [0001] The invention relates to humanized anti-tetanus exotoxin antibody (HTAT-Fab), its preparation method and application, and belongs to the field of biological products. Background technique [0002] Tetanus (tetanus) disease is an acute infectious disease characterized by muscle tonic spasm caused by tetanus bacilli invading human wounds and multiplying in anaerobic environment to produce neurotropic exotoxins. Severe patients may die due to laryngospasm or secondary severe lung infection; neonatal and severe patients, the case fatality rate can be as high as 20-50%; about 50,000 people die of tetanus every year in the world. Tetanus antitoxin (TAT) or human tetanus immunoglobulin (Human Tetanusimmunoglulin, HTIG) injections are often required in an effective tetanus prevention and treatment program. According to incomplete statistics, the domestic demand for TAT and HTIG has reached more than 10 million servings / year, and the international demand is...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P31/04C07K16/08C12N15/12
Inventor 高尚先张利宁孙汶生王群王福清刘军莉赵强
Owner 北京明新高科技发展有限公司
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