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Monitorization of alfa1-Acid Glycoprotein Deglycosylation using SU-8 Microchips Electrophoresis with LIF Detection

AuthorsBarrios-Romero, María del Mar; Crevillén, Agustín G.; Puerta, Angel de la ; Frutos, Mercedes de ; Diez-Masa, José Carlos
KeywordsMicrochip gel electrophoresis
SU-8 microchips
LIF detection
α1-acid glycoprotein
Issue Date2019
PublisherHumana Press
CitationMethods in molecular biology (Clifton, N,J,) 1972: 25-39 (2019)
AbstractIn 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 ¿1-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.
Publisher version (URL)http://dx.doi.org/10.1007/978-1-4939-9213-3_3
Identifiersdoi: 10.1007/978-1-4939-9213-3_3
issn: 1064-3745
Appears in Collections:(IQOG) Artículos
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