Please use this identifier to cite or link to this item: https://doi.org/10.1371/journal.pone.0116878
Title: Optimization of heavy chain and light chain signal peptides for high level expression of therapeutic antibodies in CHO cells
Authors: Haryadi R.
Ho S.
Kok Y.J.
Pu H.X.
Zheng L.
Pereira N.A.
Li B.
Bi X.
Goh L.-T.
Yang Y.
Song Z. 
Keywords: adalimumab
aspartic acid
aspartylyisoleucylglutaminylmethionylthreonylglutaminylserylprolylserylserylleucylserylalanylserylvalylglycylaspartylarginine
bevacizumab
cysteine
glutamic acid
glutamylserylglycylglycylglycylleucylvalylglutaminylprolylglycylglycylserylleucylarginine
glutamylvalylglutaminylleucylvalylglutamylserylglycylglycylglycylvalylglutaminylprolylglycylglycylserylleucylarginine
immunoglobulin kappa chain
immunoglobulin light chain
infliximab
methionylaspartylphenyalalyglutaminylvalylisoleucylisoleucylserylphenylalanylleucylleucylisoleucylserylalalylserylvalylisoleucylmethionylserylarginylglycine
methionylglutamylphenylalanylglycylleucylseryltryptophanylvalylphenylalanylleucylvalylalanylleucylphenylalanylarginylglycylvalylglutamylcysteine
methionylglyclytryptophanylserylleucylisoleucylleucylleucylphenylalanylleucylvalylalanylvalylalanyltyrosylarginylvalylleucylserine
peptide
rituximab
trastuzumab
unclassified drug
immunoglobulin G
immunoglobulin heavy chain
immunoglobulin light chain
monoclonal antibody
signal peptide
amino acid sequence
amino acid substitution
animal cell
antibody detection
antibody production
antibody titer
antigen expression
Article
mass spectrometry
nonhuman
peptide analysis
phylogeny
protein cleavage
protein processing
sequence alignment
sequence homology
animal
CHO cell line
Cricetulus
metabolism
physiology
Animals
Antibodies, Monoclonal
CHO Cells
Cricetulus
Immunoglobulin G
Immunoglobulin Heavy Chains
Immunoglobulin Light Chains
Mass Spectrometry
Protein Sorting Signals
Issue Date: 2015
Citation: Haryadi R., Ho S., Kok Y.J., Pu H.X., Zheng L., Pereira N.A., Li B., Bi X., Goh L.-T., Yang Y., Song Z. (2015). Optimization of heavy chain and light chain signal peptides for high level expression of therapeutic antibodies in CHO cells. PLoS ONE 10 (2) : e0116878. ScholarBank@NUS Repository. https://doi.org/10.1371/journal.pone.0116878
Rights: Attribution 4.0 International
Abstract: Translocation of a nascent protein from the cytosol into the ER mediated by its signal peptide is a critical step in protein secretion. The aim of this work was to develop a platform technology to optimize the signal peptides for high level production of therapeutic antibodies in CHO cells. A database of signal peptides from a large number of human immunoglobulin (Ig) heavy chain (HC) and kappa light chain (LC) was generated. Most of the HC signal peptides contain 19 amino acids which can be divided into three domains and the LC signal peptides contain 22 amino acids. The signal peptides were then clustered according to sequence similarity. Based on the clustering, 8 HC and 2 LC signal peptides were analyzed for their impacts on the production of 5-top selling antibody therapeutics, namely, Herceptin, Avastin, Remicade, Rituxan, and Humira. The best HC and LC signal peptides for producing these 5 antibodies were identified. The optimized signal peptides for Rituxan is 2-fold better compared to its native signal peptides which are available in the public database. Substitution of a single amino acid in the optimized HC signal peptide for Avastin reduced its production significantly. Mass spectrometry analyses revealed that all optimized signal peptides are accurately removed in the mature antibodies. The results presented in this report are particularly important for the production of these 5 antibodies as biosimilar drugs. They also have the potential to be the best signal peptides for the production of new antibodies in CHO cells. © 2015 Haryadi et al.
Source Title: PLoS ONE
URI: https://scholarbank.nus.edu.sg/handle/10635/161742
ISSN: 19326203
DOI: 10.1371/journal.pone.0116878
Rights: Attribution 4.0 International
Appears in Collections:Elements
Staff Publications

Show full item record
Files in This Item:
File Description SizeFormatAccess SettingsVersion 
10_1371_journal_pone_0116878.pdf797.64 kBAdobe PDF

OPEN

PublishedView/Download

SCOPUSTM   
Citations

62
checked on Sep 20, 2022

Page view(s)

305
checked on Sep 22, 2022

Download(s)

1
checked on Sep 22, 2022

Google ScholarTM

Check

Altmetric


This item is licensed under a Creative Commons License Creative Commons