PMID- 24498415
OWN - NLM
STAT- MEDLINE
DCOM- 20141020
LR  - 20211021
IS  - 1932-6203 (Electronic)
IS  - 1932-6203 (Linking)
VI  - 9
IP  - 2
DP  - 2014
TI  - Controllability analysis of protein glycosylation in CHO cells.
PG  - e87973
LID - 10.1371/journal.pone.0087973 [doi]
LID - e87973
AB  - To function as intended in vivo, a majority of biopharmaceuticals require 
      specific glycan distributions. However, achieving a precise glycan distribution 
      during manufacturing can be challenging because glycosylation is a non-template 
      driven cellular process, with the potential for significant uncontrolled 
      variability in glycan distributions. As important as the glycan distribution is 
      to the end-use performance of biopharmaceuticals, to date, no strategy exists for 
      controlling glycosylation on-line. However, before expending the significant 
      amount of effort and expense required to develop and implement on-line control 
      strategies to address the problem of glycosylation heterogeneity, it is 
      imperative to assess first the extent to which the very complex process of 
      glycosylation is controllable, thereby establishing what is theoretically 
      achievable prior to any experimental attempts. In this work, we present a novel 
      methodology for assessing the output controllability of glycosylation, a 
      prototypical example of an extremely high-dimensional and very non-linear system. 
      We first discuss a method for obtaining the process gain matrix for glycosylation 
      that involves performing model simulations and data analysis systematically and 
      judiciously according to a statistical design of experiments (DOE) scheme and 
      then employing Analysis of Variance (ANOVA) to determine the elements of process 
      gain matrix from the resulting simulation data. We then discuss how to use the 
      resulting high-dimensional gain matrix to assess controllability. The utility of 
      this method is demonstrated with a practical example where we assess the 
      controllability of various classes of glycans and of specific glycoforms that are 
      typically found in recombinant biologics produced with Chinese Hamster Ovary 
      (CHO) cells. In addition to providing useful insight into the extent to which 
      on-line glycosylation control is achievable in actual manufacturing processes, 
      the results also have important implications for genetically engineering cell 
      lines design for enhanced glycosylation controllability.
FAU - St Amand, Melissa M
AU  - St Amand MM
AD  - Department of Chemical and Biomolecular Engineering, University of Delaware, 
      Newark, Delaware, United States of America.
FAU - Tran, Kevin
AU  - Tran K
AD  - Department of Chemical and Biomolecular Engineering, University of Delaware, 
      Newark, Delaware, United States of America.
FAU - Radhakrishnan, Devesh
AU  - Radhakrishnan D
AD  - Department of Chemical and Biomolecular Engineering, University of Delaware, 
      Newark, Delaware, United States of America.
FAU - Robinson, Anne S
AU  - Robinson AS
AD  - Department of Chemical and Biomolecular Engineering, University of Delaware, 
      Newark, Delaware, United States of America ; Department of Chemical and 
      Biomolecular Engineering, Tulane University, New Orleans, Louisiana, United 
      States of America.
FAU - Ogunnaike, Babatunde A
AU  - Ogunnaike BA
AD  - Department of Chemical and Biomolecular Engineering, University of Delaware, 
      Newark, Delaware, United States of America.
LA  - eng
PT  - Journal Article
PT  - Research Support, U.S. Gov't, Non-P.H.S.
DEP - 20140203
PL  - United States
TA  - PLoS One
JT  - PloS one
JID - 101285081
RN  - 0 (Glycoproteins)
RN  - 0 (Polysaccharides)
SB  - IM
MH  - Animals
MH  - CHO Cells
MH  - Computer Simulation
MH  - Cricetinae
MH  - Cricetulus
MH  - Glycoproteins/*chemistry
MH  - Glycosylation
MH  - *Models, Theoretical
MH  - Polysaccharides/*analysis/*classification
PMC - PMC3912168
COIS- Competing Interests: The authors have declared that no competing interests exist.
EDAT- 2014/02/06 06:00
MHDA- 2014/10/21 06:00
PMCR- 2014/02/03
CRDT- 2014/02/06 06:00
PHST- 2013/09/23 00:00 [received]
PHST- 2014/01/02 00:00 [accepted]
PHST- 2014/02/06 06:00 [entrez]
PHST- 2014/02/06 06:00 [pubmed]
PHST- 2014/10/21 06:00 [medline]
PHST- 2014/02/03 00:00 [pmc-release]
AID - PONE-D-13-39172 [pii]
AID - 10.1371/journal.pone.0087973 [doi]
PST - epublish
SO  - PLoS One. 2014 Feb 3;9(2):e87973. doi: 10.1371/journal.pone.0087973. eCollection 
      2014.