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Preparation method of transgenic animal capable of expressing human antibody

A technology of human antibody and transgene, applied in the direction of using vectors to introduce foreign genetic material, recombinant DNA technology, animal husbandry, etc.

Active Publication Date: 2014-02-12
SHANDONG BIOANTY BIOLOGICAL TECH CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, this method can only remove the immunogenicity caused by the non-human Fc region, while the rest of the non-human Fab sequence can still be immunogenic

Method used

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  • Preparation method of transgenic animal capable of expressing human antibody
  • Preparation method of transgenic animal capable of expressing human antibody
  • Preparation method of transgenic animal capable of expressing human antibody

Examples

Experimental program
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Embodiment 1

[0053] Purification of BAC DNA Containing Human Antibody Loci

[0054] 1. Small culture of BAC bacteria. Pick a streaked single colony and inoculate it into a 15mL centrifuge tube with a culture volume of 3mL and the cap of the tube is slightly loose. 200rpm, culture at 30°C for 16-20h. The final concentration of chloramphenicol was 12.5ug / mL.

[0055] 2. Large-scale cultivation of BAC strains. Inoculate the small-cultured bacterial solution into a shaker flask at a ratio of 1:500, culture in a volume of 400 mL, 200 rpm, and incubate at 30° C. for 16-20 h. The final concentration of chloramphenicol was 12.5ug / mL.

[0056] 3. Use the MN kit to extract BAC DNA. For related steps, refer to "Endoto-free Plasmid DNA Purification User Manual--Nucleobond Xtra Midi EF". Store BAC DNA at 4°C.

[0057] 4. Use the Nanodrop to measure the concentration of the extracted BAC DNA. PFGE analysis.

[0058] 5. The purified BAC DNA can be used for microinjection directly, or after linea...

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Abstract

The invention relates to a preparation method of a mouse capable of producing a human antibody. The preparation method of the mouse capable of producing the human antibody comprises the step of carrying out hybridization on a transgenic mouse carrying a fragment containing a human antibody heavy chain gene locus part and a mouse carrying a fragment which is inserted into mouse genomic DNA (deoxyribonucleic acid) and contains an non-rearranged human antibody light chain gene locus part, wherein gene rearrangement and gene conversion can be carried out in the transgenic mouse by virtue of the human antibody heavy chain gene locus and the human antibody light chain gene locus, so as to produce various human immune globulins, and a mouse endogenous antibody heavy chain gene locus, a mouse endogenous antibody kappa light chain gene locus and a Lamda light chain gene locus are inactivated.

Description

technical field [0001] The present invention relates to a preparation method of mice producing human antibodies. Background technique [0002] Treatment of infectious diseases caused by bacteria, fungi, viruses and parasites is largely based on antibiotic therapy. However, the emergence of drug resistance requires the continuous development of new antibiotics. The treatment of patients with malignant diseases and cancers is mainly based on chemotherapy, however, many of these treatments are ineffective and the mortality rate of patients is high. For infectious diseases and cancers, improved and innovative treatments are needed. Steroidal therapy against transplanted organ rejection requires the use of biological agents (monoclonal or polyclonal antibody preparations) that reverse the ongoing alloimmune response in the transplant recipient. A major problem with antibody preparations derived from animals is the immunogenicity of non-human immunoglobulins in human patients. ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N15/85A01K67/027
Inventor 冯东晓戴洁张卉徐从伦董创创刘红
Owner SHANDONG BIOANTY BIOLOGICAL TECH CO LTD
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