Abstract:
:Chinese hamster ovary (CHO) cell lines are widely used for the expression of therapeutic recombinant proteins, including monoclonal antibodies and other biologics. For manufacturing, cells derived from a single-cell clone are typically used to ensure product consistency. Presently, fetal bovine serum (FBS) is commonly used to support low cell density cultures to obtain clonal cell populations because cells grow slowly, or even do not survive at low cell densities in protein-free media. However, regulatory authorities have discouraged the use of FBS to reduce the risk of contamination by adventitious agents from animal-derived components. In this study, we demonstrated how a complementary mass spectrometry-based shotgun proteomics strategy enabled the identification of autocrine growth factors in CHO cell-conditioned media, which has led to the development of a fully defined single-cell cloning media that is serum and animal component-free. Out of 290 secreted proteins that were identified, eight secreted growth factors were reported for the first time from CHO cell cultures. By supplementing a combination of these growth factors to protein-free basal media, single cell growth of CHO cells was improved with cloning efficiencies of up to 30%, a 2-fold improvement compared to unsupplemented basal media. Complementary effects of these autocrine growth factors with other paracrine growth factors were also demonstrated when the mixture improved cloning efficiency to 42%, similar to that for the conditioned medium.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Lim UM,Yap MG,Lim YP,Goh LT,Ng SKdoi
10.1021/pr400352nsubject
Has Abstractpub_date
2013-07-05 00:00:00pages
3496-510issue
7eissn
1535-3893issn
1535-3907journal_volume
12pub_type
杂志文章abstract::We analyzed a unique rice (Oryza sativa L.) blast lesion mimic (blm) mutant for differentially expressed proteins in leaves of one- and two-week-old seedlings manifesting the lesion mimic phenotype. Gel-based one- and two-dimensional electrophoresis (1- and 2-DGE) was performed using leaves (blm and wild-type, WT) bef...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr060092c
更新日期:2006-10-01 00:00:00
abstract::Bioactive peptides play critical roles in regulating most biological processes in animals, and have considerable biological, medical and industrial importance. A number of peptides have been discovered usually based on their biological activities in vitro or based on their sequence similarities in silico. Through sear...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr800037n
更新日期:2008-09-01 00:00:00
abstract::Roux-en-Y gastric bypass (RYGB) is one of the most effective treatments for long-term weight loss and diabetes remission; however, the mechanisms underlying these changes are not clearly understood. In this study, the serum metabolic profiles of 23 remission and 12 nonremission patients with type 2 diabetes mellitus (...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00022
更新日期:2016-04-01 00:00:00
abstract::Defined as proteins actively transported via secretion systems, secreted proteins can have radically different subcellular destinations in monoderm (Gram-positive) bacteria. From degradative enzymes in saprophytes to virulence factors in pathogens, secreted proteins are the main tools used by bacteria to interact with...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr1003642
更新日期:2010-10-01 00:00:00
abstract::The present study reports an in-depth proteome analysis of two Lactobacillus rhamnosus strains, the well-known probiotic strain GG and the dairy strain Lc705. We used GeLC-MS/MS, in which proteins are separated using 1-DE and identified using nanoLC-MS/MS, to generate high-quality protein catalogs. To maximize the num...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr2000896
更新日期:2011-08-05 00:00:00
abstract::Proteolysis of autoantigens can alter normal MHC class II antigen processing and has been implicated in the induction of autoimmune diseases. Many autoantigens are substrates for the protease granzyme B (GrB), but the mechanistic significance of this association is unknown. Peptidylarginine deiminase 4 (PAD4) is a fre...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00617
更新日期:2017-01-06 00:00:00
abstract::Prostate cancer (PCa) is the second leading cause of cancer-related mortality in men. The prevalent diagnosis method is based on the serum prostate-specific antigen (PSA) screening test, which suffers from low specificity, overdiagnosis, and overtreatment. In this work, untargeted metabolomic profiling of age-matched ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr500409q
更新日期:2014-07-03 00:00:00
abstract::Protein citrullination (or deimination), an irreversible post-translational modification, has been implicated in several physiological and pathological processes, including gene expression regulation, apoptosis, rheumatoid arthritis, and Alzheimer's disease. Several research studies have been carried out on citrullina...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.9b00118
更新日期:2019-05-03 00:00:00
abstract::An algorithm for protein identification based on mass spectrometric proteolytic peptide mapping and genome database searching is presented. The algorithm ranks database proteins based on direct calculation of the probability of random matching and assigns the statistical significance to each result. We investigate the...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr034048y
更新日期:2004-01-01 00:00:00
abstract::Plasma biomarkers of exposure to environmental contaminants play an important role in early detection of disease. The emerging field of proteomics presents an attractive opportunity for candidate biomarker discovery, as it simultaneously measures and analyzes a large number of proteins. This article presents a case st...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr070491n
更新日期:2008-01-01 00:00:00
abstract::Difference gel electrophoresis (DiGE) allows the reliable comparison of proteome differences between two or three samples within a single gel, by way of a CyDye fluorescent labeling system. This facilitates identification of protein differences avoiding the difficulties associated with gel-to-gel variation. A drawback...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr0606878
更新日期:2007-09-01 00:00:00
abstract::Stroke remains a major public health problem worldwide; it causes severe disability and is associated with high mortality rates. However, early diagnosis of stroke is difficult, and no reliable biomarkers are currently established. In this study, mass-spectrometry-based metabolomics was utilized to characterize the me...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00779
更新日期:2017-02-03 00:00:00
abstract::To investigate the role of rpoS in gene expression of Escherichia coli cells grown as biofilms, we compared the proteomes of a rpoS mutant and the wild-type strain. Experiments were performed on planktonic cells (in exponential or stationary growth phase) and biofilms developed on glass wool. Spot-by-spot comparison o...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr8001723
更新日期:2008-11-01 00:00:00
abstract::Robust quantification is an essential component of comparative -omic strategies. In this regard, glycomics lags behind proteomics. Although various isotope-tagging and direct quantification methods have recently enhanced comparative glycan analysis, a cell culture labeling strategy, that could provide for glycomics th...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr8010028
更新日期:2009-08-01 00:00:00
abstract::Mutagenesis through fast neutron (FN) radiation of soybean resulted in a mutant with a 15% increase in seed protein content. A comparative genomic hybridization analysis confirmed that the mutant is lacking 24 genes located at chromosomes 5 and 10. A tandem mass tag-based proteomic profiling of the wild type and the F...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.0c00160
更新日期:2020-10-02 00:00:00
abstract::Huntington's disease (HD) is a neurodegenerative disorder caused by expansion of a CAG repeat within the Huntingtin (HTT) gene, though the clinical presentation of disease and age-of-onset are strongly influenced by ill-defined environmental factors. We recently reported a gene-environment interaction wherein expressi...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200839d
更新日期:2012-02-03 00:00:00
abstract::Antibiotic-induced microbial perturbations alter host metabolism and affect host physiology. In this study, we aimed to investigate the effects of vancomycin (Vanc) and ciprofloxacin/metronidazole (CiMe) exposures on the gut microbiome and metabolome in the colonic content and tissue samples from advanced-stage type 1...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.9b00402
更新日期:2019-11-01 00:00:00
abstract::A well-described animal model was used to understand the molecular mechanisms of carcinogenesis and metastasis of gastric cancer at the protein level. Gastric cancer was induced in 12 Wistar rats by oral administration of N-methyl-N'-Nitro-N-Nitrosoguanidine (MNNG). The protein expression patterns of normal gastric ti...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr049916l
更新日期:2004-09-01 00:00:00
abstract::Periodontal disease is perhaps the most common infectious disease in humans. Gingival crevicular fluid (GCF) is a local inflammatory exudate of the periodontal tissues. Its composition greatly varies between health and periodontal disease. GCF collection is rapid and noninvasive, but previous approaches aiming to anal...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr900941z
更新日期:2010-05-07 00:00:00
abstract::Controlled ovarian hyperstimulation is performed to assist with generation of multiple mature oocytes for use in in vitro fertilization (IVF). The goal of our study was to evaluate differences in protein and steroid profiles in ovarian follicular fluid (hFF) samples obtained during oocyte retrieval from women undergoi...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr300535g
更新日期:2012-10-05 00:00:00
abstract::Protein biomarkers are fundamental tools for the characterization of stem cells and for tracking their differentiation and maturation down developmental lineages. Technology development allowing increased coverage of difficult cellular proteomes should allow for the discovery of new and novel membrane protein biomarke...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr900597s
更新日期:2009-12-01 00:00:00
abstract::Protein-protein interactions play a major role in the molecular machinery of life, and various techniques such as AP-MS are dedicated to their identification. However, those techniques return lists of proteins devoid of organizational structure, not detailing which proteins interact with which others. Proposing a hier...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.0c00066
更新日期:2020-07-02 00:00:00
abstract::This paper describes a metabonomic study of prenatal disorders using nuclear magnetic resonance (NMR) spectroscopy of amniotic fluid (AF) collected in the second trimester of pregnancy, to search for metabolite markers of fetal malformations, prediagnostic gestational diabetes (GD), preterm delivery (PTD), early ruptu...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr100815q
更新日期:2010-11-05 00:00:00
abstract::We report the use of chemical derivatization with MALDI-MS/MS analysis for de novo sequence analysis. Using three frequently used homology-based search algorithms, we were able to identify more than 40 proteins from banana, a nonmodel plant with unsequenced genome. Furthermore, this approach allowed the identification...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr0601943
更新日期:2007-01-01 00:00:00
abstract::The identification and the present wide acceptance of cardiovascular risk factors such as age, sex, hypertension, hyperlipidemia, smoking, obesity, diabetes, and physical inactivity have led to dramatic reductions in cardiovascular morbidity and mortality. However, novel risk predictors present opportunities to identi...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200452j
更新日期:2011-11-04 00:00:00
abstract::Linear, short-chain polyfluorinated and perfluorinated alkyl compounds, often referred to as PFCs, have been in worldwide use as surfactants and polymer precursors for decades, and environmental dispersal of these highly persistent compounds represents a public health threat. Whereas ubiquitous low-level exposure to t...
journal_title:Journal of proteome research
pub_type: 杂志文章,评审
doi:10.1021/pr500990w
更新日期:2015-01-02 00:00:00
abstract::Microglial inflammatory responses affect Parkinson's disease (PD) associated nigrostriatal degeneration. This is triggered, in measure, by misfolded, nitrated alpha-synuclein (N-alpha-syn) contained within Lewy bodies that are released from dying or dead dopaminergic neurons into the extravascular space. N-alpha-syn-s...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr9001614
更新日期:2009-07-01 00:00:00
abstract::Quantitative analysis of protein biomarkers in plasma is typically done by ELISA, but this method is limited by the availability of high-quality antibodies. An alternative approach is protein immunoprecipitation combined with multiple reaction monitoring mass spectrometry (IP-MRM). We compared IP-MRM to ELISA for the ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr400877e
更新日期:2013-12-06 00:00:00
abstract::The last two decades have seen a broad diversity of methods used to identify and/or characterize proteins in the archeological and paleontological record. Of these, mass spectrometry has opened an unprecedented window into the proteomes of the past, providing protein sequence data from long extinct animals as well as ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.7b00703
更新日期:2018-03-02 00:00:00
abstract::We present a novel straightforward method for enrichment of N-terminal peptides, utilizing charge-based fractional diagonal chromatography (ChaFRADIC). Our method is robust, easy to operate, fast, specific, and more sensitive than existing methods, enabling the differential quantitation of 1459 nonredundant N-terminal...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr400435d
更新日期:2013-09-06 00:00:00