Monitorization of α1-Acid Glycoprotein Deglycosylation Using SU-8 Microchips Electrophoresis with LIF Detection
In the last few years, biopharmaceuticals—therapeutic drugs which are generally obtained by using molecular biology techniques—have become a major growing sector in pharmaceutical industry. A large part of these biopharmaceuticals are therapeutic glycoproteins. The production of these drugs and their purification process are implying the development of efficient analytical methods, which allow quick and reliable control of the manufacturing process and ensuring the regulatory compliance about the quality of these drugs. Capillary gel electrophoresis (CGE) in the presence of sodium dodecyl sulfate (SDS) is becoming a method of choice in the quality control of these biopharmaceuticals. On the other hand, CGE can be improved if analyses are carried out in microchip format.
This chapter reports a detailed microchips gel electrophoresis (MGE) method to separate glycosylated and deglycosylated forms of α1-acid glycoprotein (AGP) labeled with Chromeo P540, using SU-8 microchips and laser induced fluorescence detection. Due to the analogy between AGP and some therapeutic glycoproteins, we have selected AGP as a model system to illustrate the potential of MGE in the analysis of this type of biopharmaceutical compounds.
Key wordsMicrochip gel electrophoresis SU-8 microchips LIF detection α1-acid glycoprotein AGP Glycoprotein Deglycosylation
The authors acknowledge the Spanish Ministries of Science and Innovation and of Economy, Industry, and Competitiveness (grants PSS-010000-2008-6 and CTQ2013-43236-R, respectively) and CSIC (joint project 2009JP003 with the Japanese Society for Promotion of Science) for financial support. M.M.B-R thanks the CSIC for a JAE-Pre contract and A.G.C. acknowledges CSIC for JAE-Doc contract. These contracts are co-financed by the European Union under the European Social Fund (ESF).
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