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Global assessment of protein turnover in recombinant antibody producing myeloma cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
24049 43492 2010 12 PDF Available
Title
Global assessment of protein turnover in recombinant antibody producing myeloma cells
Abstract

The global turnover rates of cellular proteins and the secretion rate of a recombinant immunoglobulin G (IgG) in a myeloma cell line, NS0, were determined using SILAC proteomic analysis. After complete labeling of cellular proteins with 13C6, 15N4-arginine, cells were transferred to unlabeled medium and the decay of the labeled arginine in proteins was monitored during exponential cell growth. After PAGE separation and mass-spectrometric identification of proteins, those detected with high confidence over at least three time points were used for the determination of turnover rates. Among the 224 proteins quantified with a protein half-life, about 15% have a degradation rate constant lower than one-tenth of specific growth rate. For most proteins, the turnover rate is insignificant in its overall dynamics. Only 6.3% of proteins have a half-life shorter than the cell doubling time. For IgG secretion, both heavy and light chain molecules follow the same kinetic behavior with a half-life estimated to be 2 h. The label decay curve appears to show a second region with very slow kinetics, raising the possibility of two populations of IgG molecules with different secretion characteristics.

Keywords
Proteomics; NS0; Recombinant IgG; Protein turnover; SILAC
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 148, Issue 4, 2 August 2010, Pages 182–193
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
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Full-text PDF Download
Online Support
Any Questions? feel free to contact us