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Efficient synthesis of isotopically pure isotope-coded affinity tagging reagents
Anupama Patel1, David M Perrin
1Department of Chemistry, 2036 Main Mall, University of British Columbia, Vancouver, B. C. V6T 1Z1 Canada, and PENCE Inc. (Protein Engineering Network of Centers of Excellence), 750 Heritage Medical Research Center, Edmonton, Alberta T6G-2S2 Canada.
Bioconjugate Chemistry
|January 22, 2004
Summary
A new, cost-effective synthesis of isotopically labeled ICAT linkers (isotope-coded affinity tags) offers improved yield and isotopic purity. This method enhances proteomic analysis by providing stable, heavy tags for protein quantification.
Area of Science:
- Chemical Synthesis
- Proteomics
- Analytical Chemistry
Background:
- Isotope-Coded Affinity Tags (ICAT) are crucial for quantitative proteomics.
- Existing methods for synthesizing heavy ICAT reagents face challenges in cost, yield, and isotopic retention.
Purpose of the Study:
- To develop an improved synthesis for isotopically pure heavy ICAT linkers.
- To enhance the cost-effectiveness, yield, and isotopic stability of heavy ICAT reagent production.
Main Methods:
- A seven-step synthesis was employed to create an isotopically pure d8-ICAT linker.
- Etherification of exchange-inert d4-starting materials ensured isotopic stability throughout the process.
Main Results:
- The synthesis achieved an overall yield of 33% for the d8-ICAT linker.
- The resulting synthon demonstrated excellent isotopic retention, remaining exchange-inert in subsequent reactions.
- The modified synthesis is more cost-effective and offers higher yields compared to previous methods.
Conclusions:
- This novel synthesis provides a significant advancement in the production of heavy ICAT reagents.
- The method is adaptable for incorporating additional deuterium, 13C, and/or 18O isotopes.
- The synthesis facilitates the creation of differentially encoded ICAT tags for advanced proteomic studies.