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Fusion protein

A technology of fusion protein and regulation protein, applied in the field of fusion protein, to achieve the effects of easy preparation, convenient use and simple structure

Inactive Publication Date: 2019-07-12
FOURTH MILITARY MEDICAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0002] The complement system is an important immune effect amplification system and immune regulation system in the body. It is widely involved in innate immunity and adaptive immunity. The products of complement activation have many biological functions such as lysing cells, mediating inflammation, regulating immune response and clearing immune complexes. However, complement deficiency, dysfunction or overactivation can lead to the occurrence of various diseases

Method used

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preparation example Construction

[0068] A method for preparing a fusion protein, comprising the following steps:

[0069] (1) Construction and small-scale expression of recombinant plasmids:

[0070] Design primers to construct expression vectors of prokaryotic expression vectors PET-28a (tag size about 4kD) and pCOLD-sumo (tag size about 16kD). After the construction is completed, the sequencing results are 100% correct and there are no mutations. Enter the small test for verification, and pass the dissolution aid Protocols of labeling and temperature induction achieve secreted expression. If it is still expressed as an inclusion body through multiple optimizations, it will be refolded to ensure that the final protein prepared is as correct as possible!

[0071] Basic route: primer design—vector construction—sequencing verification—transformation of expression strains (DE3 and Rosseta)—small expression verification—supernatant precipitation detection of broken bacteria

[0072] (2) Large-scale expression a...

Embodiment 1

[0084] In this example, the new fusion protein is CD55(1-356)+CD46(35-326); that is, CD55(SP+Domian1-4+ST)+CD46(Domian1-4+ST)

[0085] Gene sequence: 1068bp+876bp=1944bp

[0086] Amino acid sequence: 356aa+292aa=648aa, MW=71.175KD, pI=6.36

[0087] First, the analysis shows the fusion of this protein, then its components, then the fusion steps, and finally the embodiment of treating BP.

Embodiment 2

[0089] In this example, the new fusion protein is CD46(35-285)+CD55(96-285)+CD46(35-285); that is, CD46(Domian1-4)+CD55(Domian2-4)+CD46(Domian1- 4)

[0090] Gene sequence: 753bp+570bp+753bp=2076aa

[0091] Amino acid sequence: 251aa+190aa+251aa=692aa, MW=77.703KD, pI=5.33

[0092] First, the analysis shows the fusion of this protein, then its components, then the fusion steps, and finally the embodiment of treating BP.

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Abstract

The invention discloses a fusion protein, and belongs to the biopharmaceutical technical field. The fusion protein is a fusion protein obtained by fusion of a regulatory protein CD46 and a regulatoryprotein CD55.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and in particular relates to a fusion protein. Background technique [0002] The complement system is an important immune effect amplification system and immune regulation system in the body. It is widely involved in innate immunity and adaptive immunity. The products of complement activation have many biological functions such as lysing cells, mediating inflammation, regulating immune response and clearing immune complexes. However, complement deficiency, dysfunction or overactivation can lead to the occurrence of various diseases. Under normal physiological conditions, the body has a fine regulatory mechanism for the activation of the complement system, which keeps the complement system in a state of inhibition and no biological function, so as to maintain the stability of the internal environment. It can be activated under the stimulation of antigen-antibody complexes, microbial component...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K19/00
CPCC07K14/70596C07K2319/00
Inventor 乔佩党二乐王刚
Owner FOURTH MILITARY MEDICAL UNIVERSITY
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