Gel Electrophoretic and Mass Spectrometry-based methodologies in the quality study of Biopharmaceuticals

Session

Pharmaceutical and Natural Sciences

Description

Modern analytical methodologies play an essential role regarding the quality evaluation of complex components, such as biologicals, where their implementation turned out to be crucial to assure their quality, safety, and efficacy. Glycoprotein hormones possess a highly complex structure and the study of their quality requires advanced analytical methodologies. Therefore, the aim of this study was the evaluation of suitable analytical methods for the quality study of such complex compounds. In order to satisfy the aim, 1D-SDS-PAGE and 2D gel electrophoresis in conjunction with mass spectrometry-based analyses were implemented. Gel electrophoretic techniques revealed the pI, Mr and glycoforms pattern, while MS analyses shed light upon the identity, structural integrity and glycosylation extent. The results demonstrated high complexity and extreme heterogeneity, typical for glycoproteins. Mass spectrometry provided information regarding structural identity parameters and glycosylation model. This methodology proved to be capable for the determination of purity and structural integrity of such complex compounds, although further investigations are required to fully understand glycosylation pattern since it has a significant effect on the pharmacokinetics and their biological activity.

Keywords:

glycoproteins, glycosylation pattern, gel electrophoresis, mass spectrometry-based analyses.

Session Chair

Valon Ejupi

Session Co-Chair

Dafina Krasniqi

Proceedings Editor

Edmond Hajrizi

ISBN

978-9951-550-19-2

Location

Pristina, Kosovo

Start Date

26-10-2019 11:30 AM

End Date

26-10-2019 1:00 PM

DOI

10.33107/ubt-ic.2019.26

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Oct 26th, 11:30 AM Oct 26th, 1:00 PM

Gel Electrophoretic and Mass Spectrometry-based methodologies in the quality study of Biopharmaceuticals

Pristina, Kosovo

Modern analytical methodologies play an essential role regarding the quality evaluation of complex components, such as biologicals, where their implementation turned out to be crucial to assure their quality, safety, and efficacy. Glycoprotein hormones possess a highly complex structure and the study of their quality requires advanced analytical methodologies. Therefore, the aim of this study was the evaluation of suitable analytical methods for the quality study of such complex compounds. In order to satisfy the aim, 1D-SDS-PAGE and 2D gel electrophoresis in conjunction with mass spectrometry-based analyses were implemented. Gel electrophoretic techniques revealed the pI, Mr and glycoforms pattern, while MS analyses shed light upon the identity, structural integrity and glycosylation extent. The results demonstrated high complexity and extreme heterogeneity, typical for glycoproteins. Mass spectrometry provided information regarding structural identity parameters and glycosylation model. This methodology proved to be capable for the determination of purity and structural integrity of such complex compounds, although further investigations are required to fully understand glycosylation pattern since it has a significant effect on the pharmacokinetics and their biological activity.