Characterization of glycoprotein biopharmaceutical products by Caliper LC90 CE-SDS gel technology

Methods Mol Biol. 2013:988:199-209. doi: 10.1007/978-1-62703-327-5_12.

Abstract

Over the last decade, science has greatly improved in the area of protein sizing and characterization. Efficient high-throughput methods are now available to substitute for the traditional labor-intensive SDS-PAGE methods, which alternatively take days to analyze a very limited number of samples. Currently, PerkinElmer(®) (Caliper) has designed an automated chip-based fluorescence detection method capable of analyzing proteins in minutes with sensitivity similar to standard SDS-PAGE. Here, we describe the use and implementation of this technology to characterize and screen a large number of formulations of target glycoproteins in the 14-200 kDa molecular weight range.

MeSH terms

  • Electrophoresis, Capillary / methods
  • Electrophoresis, Polyacrylamide Gel / methods
  • Electrophoresis, Polyacrylamide Gel / standards
  • Erythropoietin / chemistry*
  • Glycoproteins / chemistry
  • Immunoglobulin Fc Fragments / chemistry*
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase / chemistry
  • Protein Array Analysis / methods
  • Protein Array Analysis / standards
  • Recombinant Fusion Proteins / chemistry
  • Reference Standards
  • Spectrometry, Fluorescence

Substances

  • Glycoproteins
  • Immunoglobulin Fc Fragments
  • Recombinant Fusion Proteins
  • Erythropoietin
  • Peptide-N4-(N-acetyl-beta-glucosaminyl) Asparagine Amidase