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A Quantitative Glycomics and Proteomics Combined Purification Strategy
Published on: March 8, 2016
A high-throughput method for quantification of glycoprotein sialylation.
Lam Raga A Markely1, Boon Tee Ong, Kong Meng Hoi
1Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Analytical Biochemistry
|August 10, 2010
Summary
A new rapid method accurately quantifies sialic acid in therapeutic glycoproteins. This high-throughput technique aids bioprocess monitoring and optimization for consistent quality.
Area of Science:
- Biochemistry
- Biotechnology
- Process Chemistry
Background:
- Sialic acid is crucial for therapeutic glycoprotein quality, affecting half-life, activity, and solubility.
- Current sialic acid quantification methods are slow and not suitable for high-throughput analysis.
- Optimized bioprocesses require rapid monitoring of sialic acid content.
Purpose of the Study:
- To develop a novel, rapid, high-throughput method for quantifying glycoprotein sialylation.
- To enable efficient process monitoring and optimization in biopharmaceutical production.
- To validate the method for monitoring sialylation in cell culture.
Main Methods:
- Chemical reduction followed by enzymatic release of sialic acid.
- Chemical derivatization for specific quantification.
- Validation using recombinant interferon gamma (IFN-γ) in CHO cell culture.
Main Results:
- The method provides accurate and specific quantification of sialic acid.
- Analysis is rapid, completing in 15 minutes.
- Requires minimal sample volume (45 μl) with a low quantitation limit (2 μM).
Conclusions:
- This novel method offers a significant advancement for high-throughput sialic acid quantification.
- It is suitable for real-time monitoring and optimization of biopharmaceutical production processes.
- The method is versatile for various upstream and downstream bioprocessing applications.
