Abstract：
:While numerous strategies exist for biomarker discovery, the bottleneck to product development and routine use at the clinic is in the verification phase of candidate biomarkers. The aim of this study was to establish a robust and high-throughput product ion monitoring (PIM) assay that is potentially capable of rapidly verifying candidates from discovery phase experiments. Using prostate-specific antigen (PSA), a model biomarker, and a routinely used mass spectrometer for discovery platforms, an ion trap (LTQ, Thermo), the utility of this instrument to perform PIM was explored. The proteotypic doubly charged intact peptide LSEPAELTDAVK ( m/ z 637) fragmenting to m/ z 943 (PAELTDAVK) was monitored. A limit of detection of 10 attomoles with a coefficient of variation (CV) of <20% was obtained for a purified recombinant PSA digest. Immunoextraction of endogenous PSA from serum using a monoclonal antibody on a 96-well microtiter plate, followed by PIM on the LTQ, enabled quantification of PSA down to less than 1 ng/mL with a limit of detection of 0.1 ng/mL and CVs < 20%. Mascot searching and ion ratio confirmation further supported the conclusion that the quantified moiety in serum was the PSA peptide. We conclude that this methodology could be adapted quickly and easily to other candidates, thus providing a much needed technology to bridge the gap between discovery and validation platforms.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Kulasingam V,Smith CR,Batruch I,Buckler A,Jeffery DA,Diamandis EPdoi
10.1021/pr7005999subject
Has Abstractpub_date
2008-02-01 00:00:00pages
640-7issue
2eissn
1535-3893issn
1535-3907journal_volume
7pub_type
杂志文章abstract：:The current technique used for microbial identification in hospitals is matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). However, it suffers from important limitations, in particular, for closely related species or when the database used for the identification lacks the appr...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00351
更新日期：2021-01-01 00:00:00
abstract：:Recent studies have shown a remarkable degree of plasticity in the N-glycome of the model nematode Caenorhabditis elegans; ablation of glycosylation-relevant genes can result in radically altered N-glycan profiles despite only minor biological phenotypic effects. Up to four fucose residues and five different linkages ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00746
更新日期：2015-12-04 00:00:00
abstract：:Peptide abundance estimation is generally the first step in protein quantification. In peptide abundance estimation, peptide overlapping and peak intensity variation are two challenges. The main objective of this paper is to estimate peptide abundance by taking advantage of peptide isotopic distribution and smoothness...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100181g
更新日期：2010-05-07 00:00:00
abstract：:Many proteins can be modified by multiple types of post-translational modifications (Mtp-proteins). Although some post-translational modifications (PTMs) have recently been found to be associated with life-threatening diseases like cancers and neurodegenerative disorders, the underlying mechanisms remain enigmatic to ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr401019d
更新日期：2014-06-06 00:00:00
abstract：:Multiple reaction monitoring (MRM) is emerging as a promising technique for the detection and quantification of protein biomarkers in complex biological samples. Compared to Western blotting or enzyme assays, its high sensitivity, specificity, accuracy, assay speed, and sample throughput represent a clear advantage fo...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr4006712
更新日期：2013-12-06 00:00:00
abstract：:Large-scale identification of protein-protein interactions (PPIs) in functional complexes represents an efficient route to elucidate the regulatory rules of cellular functions. Whereas many methods have been developed to identify the PPIs associated with particular target/bait protein in complexes, little information ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800662x
更新日期：2009-05-01 00:00:00
abstract：:We used a peptidomic approach for the analysis of the low molecular weight proteome in rat pancreatic tissue extracts. The goal was to develop a method that allows identifying endogenous peptides produced in the pancreas in the course of acute pancreatitis. The workflow combines peptides enrichment by centrifugal ultr...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr1002627
更新日期：2010-09-03 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：:Huntington's disease (HD) causes widespread CNS changes and systemic abnormalities including endocrine and immune dysfunction. HD biomarkers are needed to power clinical trials of potential treatments. We used multiplatform proteomic profiling to reveal plasma changes with HD progression. Proteins of interest were eva...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0700753
更新日期：2007-07-01 00:00:00
abstract：:We recently developed a novel method for analysis of collagen O-glycosylations, which include galactosyl-hydroxylysine (GHL) and glucosyl-galactosyl-hydroxylysine (GGHL), using hydrazide chemistry (Taga, Y., Mol. Cell. Proteomics 2012, 11 (6), M111.010397). Here we investigated an overglycosylation model of collagen p...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr400079d
更新日期：2013-05-03 00:00:00
abstract：:Nanobacteria may cause peripheral neuropathy by adhesion to the perineurium. This hypothesis receives support from five independent observations: (1) identification of perineurial apatite in diabetic patients with peripheral neuropathy, (2) massive presence of nanobacteria in a diabetic patient, (3) beneficial effect ...
journal_title：Journal of proteome research
pub_type： 信件
doi：10.1021/pr049935v
更新日期：2004-09-01 00:00:00
abstract：:N-Glycans are an important source of milk oligosaccharides. In addition to free oligosaccharides found in milk, N-glycans can also be utilized by gut microbes. A potential for milk N-glycans to act as gut microbe regulators in suckling animals has attracted considerable attention; however, sow milk N-glycans and their...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.8b00789
更新日期：2019-03-01 00:00:00
abstract：:Activation of AMP-activated protein kinase (AMPK) upon energy depletion stimulates energy production and limits energy utilization. Erythrocytes lacking AMPK are susceptible to suicidal cell death (eryptosis). A hallmark of eryptosis is cell membrane scrambling with phosphatidylserine exposure at the erythrocyte surfa...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr101004j
更新日期：2011-04-01 00:00:00
abstract：:Completion of the human genome sequence has provided scientists with powerful resources with which to explore the molecular events associated with disease states such as diabetes. Understanding the relative levels of expression of gene products, especially of proteins, and their post-translational modifications will b...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr050024a
更新日期：2005-05-01 00:00:00
abstract：:New mass-tagging reagents for quantitative proteomics measurements have been designed using solid phase peptide synthesis technology. The solid phase mass tags have been used to accurately measure the relative amounts of cysteine-containing peptides in model peptide mixtures as well as in mixtures of tryptic digests i...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr034081k
更新日期：2004-01-01 00:00:00
abstract：:Triply and doubly charged iTRAQ ( isobaric tagging for relative and absolute quantitation) labeled peptide cations from a tryptic peptide mixture of bovine carbonic anhydrase II were subjected to electron transfer ion/ion reactions to investigate the effect of charge bearing modifications associated with iTRAQ on the ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr8001113
更新日期：2008-09-01 00:00:00
abstract：:Mitosis is a highly regulated process in which errors can lead to genomic instability, a hallmark of cancer. During this phase of the cell cycle, transcription is silent and RNA translation is inhibited. Thus, mitosis is largely driven by post-translational modification of proteins, including phosphorylation, methylat...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100281h
更新日期：2010-09-03 00:00:00
abstract：:The nation-wide electronic health record database acts as an interoperable repository of health data obtained throughout citizen contacts with health care providers. This system improves accessibility for citizens and researchers to health data with the ability to assign context to disease development. In that system,...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr1007784
更新日期：2011-01-07 00:00:00
abstract：:Porcine reproductive and respiratory syndrome virus (PRRSV) is a major threat to the swine industry worldwide and hence global food security, exacerbated by a newly emerged highly pathogenic (HP-PRRSV) strain from China. PRRSV nonstructural protein 2 (nsp2) is a multifunctional polypeptide with strain-dependent influe...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00396
更新日期：2016-05-06 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：:Gas chromatography-mass spectrometry (GC-MS) platforms are typically run in electron ionization (EI) mode for mass spectral matching and metabolite annotation. With the advent of high resolution mass spectrometry (HRMS), soft ionization techniques such as chemical ionization (CI) may provide additional coverage for co...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00774
更新日期：2020-07-02 00:00:00
abstract：:An individually darkened leaf model was used to study protein changes in the Arabidopsis mutant stay-green1 (sgr1) to partially mimic the process of leaf covering senescence that occurs naturally in the shaded rosettes of Arabidopsis plants. Utilizing this controlled and predictable induced senescence model has allowe...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300691k
更新日期：2012-11-02 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：:The plasma of patients with stable carotid atherosclerosis (n = 9), and healthy subjects (n = 10) have been fingerprinted with both GC-MS and (1)H NMR. Principal component analysis (PCA), partial least-squares-discriminant analysis (PLS-DA) and orthogonal partial least-squares-discriminant analysis (OPLS-DA) have been...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900668v
更新日期：2009-12-01 00:00:00
abstract：:Major depressive disorder (MDD) is a debilitating mental disease with a pronounced impact on the quality of life of many people; however, it is still difficult to diagnose MDD accurately. In this study, a nontargeted metabolomics approach based on ultra-high-performance liquid chromatography equipped with quadrupole t...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.5b00144
更新日期：2015-05-01 00:00:00
abstract：:Proteomics data analysis strongly benefits from not studying single proteins in isolation but taking their multivariate interdependence into account. We introduce PerseusNet, the new Perseus network module for the biological analysis of proteomics data. Proteomics is commonly used to generate networks, e.g., with affi...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.8b00927
更新日期：2019-05-03 00:00:00
abstract：:Hypoxia inducible factor (HIF)-2α protein expression in solid tumors promotes stem-like phenotype in cancer stem cells and increases tumorigenic potential in nonstem cancer cells. Recently, we have shown that HIF-1/2α gene expression is correlated to neuroblastoma (NB) poor survival and to undifferentiated tumor state...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00457
更新日期：2016-10-07 00:00:00
abstract：:In the analysis of proteins in complex samples, pre-fractionation is imperative to obtain the necessary depth in the number of reliable protein identifications by mass spectrometry. Here we explore isoelectric focusing of peptides (peptide IEF) as an effective fractionation step that at the same time provides the adde...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0601180
更新日期：2006-07-01 00:00:00
abstract：:We have developed herein a quantitative mass spectrometry-based approach to analyze the etiology-related alterations in fucosylation degree of serum haptoglobin in patients with liver cirrhosis and hepatocellular carcinoma (HCC). The three most common etiologies, including infection with hepatitis B virus (HBV), infec...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr500128t
更新日期：2014-06-06 00:00:00
abstract：:Human milk oligosaccharides (HMOs) perform a number of functions including serving as prebiotics to stimulate the growth of beneficial intestinal bacteria, as receptor analogues to inhibit binding of pathogens, and as substances that promote postnatal brain development. There is further evidence that HMOs participate ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100362f
更新日期：2010-08-06 00:00:00
