CKMB Human S.Chain
Description
Introduction to CKMB Human Single Chain
Creatine Kinase MB Human Single Chain (CKMB Human S.Chain) is a recombinant form of the CK-MB isoenzyme, a critical biomarker for myocardial injury. Unlike native CK-MB, which exists as a heterodimer of M and B subunits (84 kDa total molecular weight), the single-chain variant is engineered as a unified polypeptide chain (~44 kDa) expressed in Escherichia coli . This innovation simplifies production while retaining diagnostic utility in cardiac pathology and research applications.
Production and Purification
Recombinant CKMB Human S.Chain is synthesized using advanced tandem expression systems. Key methodologies include:
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Gene Cloning: M and B subunit genes from human cardiac cDNA libraries are co-expressed in E. coli .
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Purification: Proprietary chromatographic techniques ensure >90% purity (SDS-PAGE) .
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Tagging Strategies: Dual affinity tags (e.g., His-tag and GST) enable efficient separation and characterization of subunits .
Table 2: Activity Comparison
| CK-MB Type | Specific Activity (U/mg) | Source |
|---|---|---|
| Native Human Heart | >25 | |
| Recombinant S.Chain | >275 | |
| Yeast-Derived | ~25 |
Clinical Use
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Acute Myocardial Infarction (AMI): CKMB Human S.Chain serves as a calibrator in mass assays (e.g., Siemens Healthineers’ Acute Care™ CK-MB test), achieving results in 14 minutes .
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Cancer Biomarker: Elevated CK-MB/CK ratios (>1.238) correlate with malignancies, aiding early detection .
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Neurodegenerative Diseases: Elevated serum CK-MB levels in ALS patients (53.85% positivity) suggest neuromuscular involvement .
Standardization
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Assay Calibration: Recombinant CK-MB reduces inter-assay bias by 15–59% compared to native standards .
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Matrix Compatibility: Stripped human serum optimizes recovery (60–100%) in immunoassays .
Prognostic Value in Stable Coronary Disease
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Mortality Prediction: High-sensitivity CK-MB (hsCK-MB ≥4.73 ng/mL) independently predicts all-cause mortality (HR 2.08, p <0.0026) but not cardiovascular events .
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Multi-Biomarker Limitations: Combining hsCK-MB with NT-proBNP or H-FABP fails to improve prognostic accuracy .
Longitudinal Stability
Product Specs
Creatine Kinase (CK) exists in three isoenzyme forms: MM, MB, and BB, distributed in muscle, cardiac, and brain tissues, respectively. These recombinant proteins are valuable tools for calibrating diagnostic instruments and investigating neuromuscular diseases. Creatine Kinases have implications for various neuromuscular applications, including but not limited to cardiac disease, mitochondrial disorders, inflammatory myopathies (e.g., myasthenia gravis, polymyositis), McArdle's disease, neuromuscular junction disorders, muscular dystrophy, amyotrophic lateral sclerosis (ALS), thyroid disorders (hypothyroidism and hyperthyroidism), central core disease, acid maltase deficiency, myoglobinuria, rhabdomyolysis, motor neuron diseases, and rheumatic diseases. These conditions often present with altered Creatine Kinase levels, either elevated or reduced.
Recombinant Human Creatine Kinase MB Single Chain, expressed in E. coli, is a non-glycosylated polypeptide chain with a molecular weight of approximately 44 kDa. It is purified using proprietary chromatographic methods.
Sterile Filtered colorless liquid.
The protein is supplied in a solution containing 50% glycerol.
CKMB is stable at 10°C for 7 days but should be stored at or below -18°C for long-term storage. Avoid repeated freeze-thaw cycles.
Purity is determined to be greater than 95.0% by SDS-PAGE analysis.
Escherichia Coli.
Product Science Overview
Creatine kinase (CK) is an enzyme that plays a crucial role in cellular energy homeostasis. It catalyzes the reversible transfer of a phosphate group from ATP to creatine, forming phosphocreatine and ADP. This reaction is vital for tissues with high and fluctuating energy demands, such as muscle and brain tissues .
Recombinant human CK-MB is produced using Escherichia coli (E. coli) expression systems. This recombinant form is a single, non-glycosylated polypeptide chain with a molecular weight of approximately 44 kDa . The production involves inserting the gene coding for CK-MB into E. coli, which then expresses the protein. The protein is subsequently purified using chromatographic techniques to achieve high purity levels .
Recombinant CK-MB is used extensively in clinical laboratories for calibrating diagnostic instruments and as a reference material. It is also employed in research related to neuromuscular diseases and cardiac conditions. The stability and high purity of recombinant CK-MB make it an ideal standard for various assays .
The production of recombinant CK-MB involves several steps:
- Gene Synthesis and Cloning: The genes coding for CK-M and CK-B are synthesized and combined with tags for tandem affinity purification (TAP). These are then inserted into a plasmid vector and transformed into E. coli .
- Expression: The transformed E. coli cells express the CK-MB protein, which is then harvested.
- Purification: The protein is purified using TAP, which involves the use of tags to isolate the protein from other cellular components. This method ensures high purity and stability of the recombinant protein .
