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Human creatine kinase single-chain variable fragment with molecular chaperone function

A creatine kinase, single-chain antibody technology, applied in the direction of antibodies, anti-enzyme immunoglobulins, and the introduction of foreign genetic material using vectors, can solve the problems of large side effects and low efficacy.

Active Publication Date: 2012-05-30
BEIJING NORMAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

When using this type of molecular chaperones as a treatment for folding diseases, problems such as low curative effect and large side effects will be encountered

Method used

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  • Human creatine kinase single-chain variable fragment with molecular chaperone function
  • Human creatine kinase single-chain variable fragment with molecular chaperone function
  • Human creatine kinase single-chain variable fragment with molecular chaperone function

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Embodiment Construction

[0028] 1. Experimental materials

[0029] The host strain E.coli BL21 of the recombinant HMCK plasmid Pet-21b was preserved by the Department of Biochemistry of Beijing Normal University; the Escherichia coli strain XLI-Blue, the Escherichia coli strain HB2151 without amber suppressor, VCSM13 and the large-capacity phage antibody library were preserved by the Naval General Hospital Stored in the experimental department of the central laboratory; restriction endonucleases are products of TaKaRa Company; ovalbumin (OA) and ferritin (Fer) are purchased from Sigma Company; various enzyme-labeled antibodies are purchased from Promega Company and Sigma Company; antigen tube (Immunotube) Purchased from NUNC Company of Denmark; β-mercaptoethanol, guanidine hydrochloride (Gdn-HCl) and SDS were from Merck; ATP, acrylamide and methylene bisacrylamide were all Fluka products; Sepharose Fast Flow, CL-6B gel, HisTrap series GE products; MOPS, IPTG, AMP, and Gly are Amresco products; other r...

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Abstract

The invention discloses human CKSCFV (creatine kinase single-chain variable fragment) with a molecular chaperone function, which comprises two amino acid sequences. The invention further provides a gene sequence, a carrier and a host cell of the human CKSCFV, and specifically describes the operation process. The human CKSCFV can be interacted with human creatine kinase to facilitate correct folding of the human creatine kinase, and fine effects are achieved. The CKSCFV has fine application prospect and is applicable to the production process of human creatine kinase drug additives.

Description

technical field [0001] The present invention relates to the preparation of antibodies. Specifically, the present invention discloses two single-chain antibodies that are molecular chaperones of human creatine kinase. The invention also relates to the preparation process of the molecular chaperone of human creatine kinase. The single-chain antibody of the invention has the function of promoting the correct folding of creatine kinase, and the antibody of the invention can be used in the production process of human creatine kinase medicine, and has certain application value in the field of pharmacy. Background technique [0002] Creatine Kinase (Creatine Kinase, CK) usually exists in the cytoplasm and mitochondria of animal muscle, heart, brain and other tissues, and is an important kinase directly related to intracellular energy transfer, muscle contraction, and ATP regeneration [Kuby S , Noltman E.Creatine kinase.The Enzymes,1962,6:515-6030.Alan C,Mcluaghin.The interaction ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C07K16/40C12N15/13C12N15/63C12N1/15C12N1/19C12N1/21C12N5/10A61K39/395
Inventor 李森
Owner BEIJING NORMAL UNIVERSITY
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