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A method for efficient isotopic labeling of recombinant proteins.
1Department of Biochemistry, Weill Medical College of Cornell University, New York, NY 10021, USA.
Journal of Biomolecular NMR
|June 30, 2001
Summary
This study presents a cost-effective method for producing isotopically labeled proteins. The approach significantly reduces isotope costs for applications in structural biology and proteomics research.
Area of Science:
- Biochemistry
- Molecular Biology
- Proteomics
Background:
- Isotopically labeled proteins are crucial for structural and functional studies.
- Current methods for protein labeling can be expensive and time-consuming.
Purpose of the Study:
- To develop a rapid, efficient, and cost-effective method for preparing isotopically labeled recombinant proteins.
- To demonstrate the utility of this method for various proteins and labeling schemes.
Main Methods:
- A novel cell mass production and media exchange strategy was employed.
- High cell density cultures were used with a short incubation in labeled media.
- Protein expression was induced after metabolite clearance and growth recovery.
Main Results:
- The method was successfully applied to label angiopoietin-2 (13C), ubiquitin (15N), and E. coli lipoprotein Lpp-56 (2H/13C/15N).
- Expression yields were comparable to standard methods.
- A fourfold to eightfold reduction in isotope costs was achieved.
Conclusions:
- This approach offers a significant cost reduction for isotopic labeling of recombinant proteins.
- The method is versatile and applicable to various proteins and labeling requirements.
- It provides a practical solution for researchers needing labeled proteins for structural and functional analyses.