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A method for preparing transgenic animals capable of expressing human antibodies

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

Active Publication Date: 2016-11-23
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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  • A method for preparing transgenic animals capable of expressing human antibodies
  • A method for preparing transgenic animals capable of expressing human antibodies
  • A method for preparing transgenic animals capable of expressing human antibodies

Examples

Experimental program
Comparison scheme
Effect test

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 present invention relates to a method for preparing a mouse capable of producing human antibodies, the method comprising combining a transgenic mouse carrying a partial fragment of a heavy chain locus of a human antibody, and a human antibody carrying an unrearranged human antibody inserted into the genomic DNA of the mouse. Mouse crosses of partial fragments of antibody light chain loci that can undergo gene rearrangement and gene conversion in transgenic mice to produce various human immunoglobulins. Among them, the mouse endogenous antibody heavy chain locus, the mouse endogenous antibody κ light chain locus and the Lamda light chain locus were 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 Patents(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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