CK-MB type creatine kinase isozyme and preparation method and application thereof

A creatine kinase and isoenzyme technology, applied in the field of enzyme engineering, can solve the problems of high price and hindering the research and development of CK-MB type creatine kinase isoenzyme diagnostic products, etc.

Inactive Publication Date: 2020-03-24
杭州博茵生物技术有限公司
View PDF8 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the limited source and high price, it seriously hinders the research and development of CK-MB creatine kinase isoenzyme diagnostic products

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • CK-MB type creatine kinase isozyme and preparation method and application thereof
  • CK-MB type creatine kinase isozyme and preparation method and application thereof
  • CK-MB type creatine kinase isozyme and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment Construction

[0047] The present invention is described in further detail with reference to the accompanying drawings and specific embodiments.

[0048] 1. Construction of Creatine Kinase M Subunit and B Subunit Expression Vectors

[0049] Using OptimumGene TM The gene design software optimizes the coding gene sequence of the two subunits of humanized creatine kinase isoenzyme, replaces the rare codons of Escherichia coli, balances the GC content, and reduces the stable secondary structure near the mRNA start codon. The optimized gene was provided by Nanjing Synthesized by GenScript Biotechnology Co., Ltd. (SEQ ID NO.1, SEQ ID NO.2);

[0050]Using SEQ ID NO.1 as a template, PCR amplification was carried out with SEQ ID NO.3 and SEQ ID NO.4 as primers, and the M subunit gene fragment was recovered by gel purification; using SEQ ID NO.2 as a template, using SEQ ID NO. NO.5 and SEQ ID NO.6 were used as primers for PCR amplification, gel purification and recovery to obtain the B subunit gene ...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

PUM

PropertyMeasurementUnit
molecular weightaaaaaaaaaa
Login to view more

Abstract

The invention belongs to the technical field of enzyme engineering, and particularly relates to a CK-MB type creatine kinase isozyme and a preparation method and application thereof. The CK-MB type creatine kinase isozyme obtained by the preparation method provided by the invention is relatively high in yield, and the yield of M subunit protein and B subunit protein after equal-proportion renaturation is 35mg/L. The CK-MB type creatine kinase isoenzyme obtained by the preparation method disclosed by the invention performs quantitative detection by using a Roche creatine kinase isoenzyme detection kit (an electrochemical luminescence method), and the activity rate of the CK-MB type creatine kinase isoenzyme is greater than 41%. The preparation method of the CK-MB type creatine kinase isoenzyme provided by the invention is simple to operate, low in cost and high in yield. The prepared CK-MB type creatine kinase isoenzyme is high in biological activity and good in stability, can be used as a quality control product or a calibration product in in-vitro diagnosis, and can also be used as immunogen or screening antigen for creatine kinase isoenzyme antibody development.

Description

technical field [0001] The invention belongs to the technical field of enzyme engineering, and in particular relates to a CK-MB type creatine kinase isoenzyme, a preparation method and an application. Background technique [0002] Creatine kinase isoenzyme is a dimeric enzyme, widely present in various tissues, with a molecular weight of 81-82KDa, composed of two identical or similar subunits, including muscle type (CK-MM), brain type ( CK-BB), hybrid type (CK-MB), and mitochondrial type (MiMi). Among them, CK-MB type creatine kinase isoenzyme has the highest diagnostic specificity, and its content rises to twice the normal concentration 6 hours after acute myocardial infarction (AMI), reaches the peak after 24 hours, and drops to the baseline level after 36 hours. When a stroke occurs, the concentration of serum CK-MB can increase to 10-25 times the normal level, exceeding 10-12 times the total activity of CK. Clinically, CK-MB is regarded as the "gold standard" for detect...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to view more

Application Information

Patent Timeline
no application Login to view more
Patent Type & Authority Applications(China)
IPC IPC(8): C12N9/12C12N15/70C12Q1/48C07K16/40
CPCC07K16/40C12N9/1223C12Q1/485C12Y207/03002
Inventor 邓春泉吴配配李因来周旭一
Owner 杭州博茵生物技术有限公司
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Try Eureka
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap