• español
    • English
  • Login
  • English 
    • español
    • English

UniversidaddeCádiz

Área de Biblioteca, Archivo y Publicaciones
Communities and Collections
View Item 
  •   RODIN Home
  • Institutos de Investigación
  • Instituto de Investigación en Biomoléculas INBIO
  • Artículos Científicos INBIO
  • View Item
  •   RODIN Home
  • Institutos de Investigación
  • Instituto de Investigación en Biomoléculas INBIO
  • Artículos Científicos INBIO
  • View Item
JavaScript is disabled for your browser. Some features of this site may not work without it.

N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches

Thumbnail
Identificadores

URI: http://hdl.handle.net/10498/20379

DOI: 10.1016/j.jprot.2015.04.012

Files
2015-N-glycosylation profile analysis of Trastuzumab biosimilar candidates by Normal Phase Liquid Chromatography and MALDI-TOF MS approaches.pdf (1.093Mb)
Statistics
View statistics
Metrics and citations
 
Share
Export
Export reference to MendeleyRefworksEndNoteBibTexRIS
Metadata
Show full item record
Author/s
Sanchez-De Melo, Ivan; Grassi, Paola; Ochoa, Francisco; Bolívar Pérez, JorgeAuthority UCA; García Cózar, Francisco JoséAuthority UCA; Durán Ruiz, María del CarmenAuthority UCA
Date
2015-04
Department
Bioquímica y Biología Molecular, Microbiología, Medicina Preventiva, Salud Pública
Source
JOURNAL OF PROTEOMICS 127 (2015) 225 – 233
Abstract
The pharmaceutical market has entered an era in which the production of new therapeutics is being often replaced by “biosimilars”, copies of already commercialized products waiting for the patents to expire in order to be distributed in a more competitive and affordable manners. Due to its relevance, the ErbB2-targeted monoclonal antibody Trastuzumab (Herceptin) used as breast cancer therapy is one of the main targets in the production of biosimilars. A major challenge is to produce antibodies with the same or the closest N-glycosylation pattern seen in the commercialized drug. Several factors, such as growing conditions or cell types employed, can determine the final composition and structure of the glycans, significantly affecting the properties of the generated antibodies. Therefore, an appropriate characterization is essential. In the present study, we describe two different but complementary strategies to characterize the N-glycosylation of two biosimilar candidates of Trastuzumab. In the first case, N-glycans are fluorescently labeled and separated by Normal Phase HPLC. Different sugars will elute at different times and can be identified using specific oligosaccharide standards. In the second approach, released glycans are permethylated and analyzed by MALDI-TOF MS, being able to determine the structure because of the differential sugar masses. Biological significance The characterization of the N-glycosylation sites of therapeutic recombinant monoclonal antibodies (mAbs) is usually one of the most critical and time consuming steps in the developing process of biosimilars or any other glycosylated drug. Herein we describe two different but complementary approaches to characterize mAbs glycosylation patterns, the use of glycan fluorescence labeling coupled to HPLC and MALDI-TOF MS profile analysis. This article is part of a Special Issue entitled: HUPO 2014.
Subjects
Monoclonal antibodies; Biosimilars; N-glycosylation; MALDI-TOF MS; Normal Phase Liquid; Chromatography; 2AB fluorescent labeling, HILIC
Collections
  • Artículos Científicos [4216]
  • Articulos Científicos Biomedicina [171]
  • Artículos Científicos INBIO [242]
Attribution-NonCommercial-NoDerivatives 4.0 Internacional
This work is under a Creative Commons License Attribution-NonCommercial-NoDerivatives 4.0 Internacional

Browse

All of RODINCommunities and CollectionsBy Issue DateAuthorsTitlesSubjectsThis CollectionBy Issue DateAuthorsTitlesSubjects

My Account

LoginRegister

Statistics

View Usage Statistics

Información adicional

AboutDeposit in RODINPoliciesGuidelinesRightsLinksStatisticsNewsFrequently Asked Questions

RODIN is available through

OpenAIREOAIsterRecolectaHispanaEuropeanaBaseDARTOATDGoogle Academic

Related links

Sherpa/RomeoDulcineaROAROpenDOARCreative CommonsORCID

RODIN está gestionado por el Área de Biblioteca, Archivo y Publicaciones de la Universidad de Cádiz

Contact informationSuggestionsUser Support