Assessment of stable isotope incorporation into recombinant proteins

作者：Zhang Xin^*; Luo Quanzhou; Apostol Izydor; Luo Shun; Jerums Matthew; Huang Gang; Jiang Xinzhao Grace; Gastwirt Jessica; Savjani Nimesh; Lewis Jeffrey; Keener Ronald III; Wypych Jette

来源：Analytical Biochemistry, 2013, 433(2): 137-149.

DOI：10.1016/j.ab.2012.10.013

• Stable isotope labeling
• Mass spectrometry
• Quantitative assessment of stable isotope incorporation

Stable isotope labeling combined with mass spectrometry has been widely used in a diverse set of applications in the biochemistry and biomedical fields. When stable isotope-labeled proteins are produced via metabolic labeling of cell culture, a comprehensive assessment of the labeling pattern is imperative. In this study, we present a set of mass spectrometry-based bioanalytical tools developed for quantitatively tracing the levels of the stable isotopes incorporated into the recombinant proteins (monoclonal antibodies and Fc fusion proteins expressed in different host systems) that include total mass analysis, peptide mapping analysis, and amino acid analysis. We show that these three mass spectrometry-based analytical methods have distinctive advantages and limitations and that they are mutually complementary in evaluating the quality of stable isotope-labeled proteins. In addition, we show that the analytical techniques developed here are powerful tools to provide valuable insights into studying cell metabolism and performing flux analysis during cell culture.

• 出版日期2013-2-15