2-DE and MALDI-TOF-MS analysis of therapeutic fusion protein abatacept

Electrophoresis. 2011 Jun;32(12):1438-43. doi: 10.1002/elps.201100029. Epub 2011 May 30.

Abstract

2-DE and MALDI-TOF MS are useful techniques for the quality evaluation of medicinal products derived from recombinant DNA technology. The principal objective of this study has been to evaluate the suitability of 2-DE in combination with MALDI-TOF MS for the quality study of the therapeutic recombinant protein, abatacept. 1-DE SDS-PAGE, under reducing and nonreducing conditions, and 2-DE analysis were used for the assessment of M(r) , pI, and enzymatic deglycosylation efficiency of abatacept. 2-DE allowed the assessment of product identity, purity, charge heterogeneity, isoform pattern, and post-translational modifications. Furthermore, optimization of the deglycosylation procedure, charge heterogeneity, and sample preparation for the subsequent MALDI-TOF MS analysis has been addressed. PMF analysis allowed rapid identity confirmation of abatacept.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Abatacept
  • Amino Acid Sequence
  • Databases, Factual
  • Electrophoresis, Gel, Two-Dimensional / methods*
  • Glycosylation
  • Immunoconjugates / chemistry*
  • Immunoconjugates / metabolism
  • Molecular Sequence Data
  • Recombinant Fusion Proteins / chemistry*
  • Recombinant Fusion Proteins / metabolism
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Immunoconjugates
  • Recombinant Fusion Proteins
  • Abatacept