An enoxaparin reference standard was purchased from SANDOZ. 2-Aminoacridone (AMAC) and sodium cyanoborohydride were purchased from Sigma-Aldrich (St. Louis, MO, USA). Three 13C-labeled 8-mer calibrants were synthesized using the chemoenzymatic synthesis method.10 (link) Recombinant heparan lyase I, II, and III were expressed in E. coli and purified by a Ni-agarose column. DEAE-Sepharose Fast Flow resin was purchased from GE Healthcare (Chicago, IL, USA). All reagents and chemicals were high-performance LC (HPLC) grade or LC-mass spectrum (MS) grade.
Protocol detail
Chemoenzymatic Synthesis of 13C-labeled Heparan Sulfate
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2 aminoacridone
Deae
E coli
Enoxaparin
Lyase
Mass spectrum
Resin
Sepharose
Sodium cyanoborohydride
Synthesis
Corresponding Organization : University of North Carolina at Chapel Hill
Other organizations : Loyola University Chicago, Loyola University Medical Center, Emory University
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Variable analysis
independent variables
- Chemoenzymatic synthesis method used to synthesize three 13C-labeled 8-mer calibrants
- Not explicitly mentioned
- Enoxaparin reference standard purchased from SANDOZ
- 2-Aminoacridone (AMAC) and sodium cyanoborohydride purchased from Sigma-Aldrich
- Recombinant heparan lyase I, II, and III expressed in E. coli and purified by a Ni-agarose column
- DEAE-Sepharose Fast Flow resin purchased from GE Healthcare
- All reagents and chemicals were high-performance LC (HPLC) grade or LC-mass spectrum (MS) grade
- Positive control: Not specified
- Negative control: Not specified
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