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：:A robust method has been developed that allows analysis of both N- and O-linked oligosaccharides released from glycoproteins separated using 2D-PAGE and then electroblotted to PVDF membrane. This analysis provides efficient oligosaccharide profiling applicable to glycoproteomic analysis. The method involves the enzyma...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr025538d
更新日期：2002-11-01 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：:Ovarian carcinoma is the most lethal gynecological malignancy in worldwide. The discovery of reliable marker for early detection of ovarian carcinoma is critical for increasing patient's survival because high mortality rate is associated with late diagnosis of this tumor. In the present study, we performed comparative...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr900811x
更新日期：2010-01-01 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：:A rate-limiting and costly step in many proteomics analyses is the cloning of all of the ORFs for an organism into technique-specific vectors. Here, we describe the generation of a Campylobacter jejuni expression clone set using a high-throughput cloning approach based on recombination in E. coli. The approach uses na...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0341134
更新日期：2004-05-01 00:00:00
abstract：:One of the ultimate goals of systems biology research is to obtain a comprehensive understanding of the control mechanisms of complex cellular metabolisms. Metabolic Flux Analysis (MFA) is a important method for the quantitative estimation of intracellular metabolic flows through metabolic pathways and the elucidation...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr2002885
更新日期：2011-08-05 00:00:00
abstract：:Influenza viruses continue evolving and have the ability to cause a global pandemic, so it is very important to elucidate its pathogenesis and find new treatment methods. In recent years, proteomics has made important contributions to describing the dynamic interaction between influenza viruses and their hosts, especi...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00778
更新日期：2021-01-01 00:00:00
abstract：:Ubiquitin ligases (E3s) serve as key regulators for the ubiquitylation-mediated pathway. The identification of the corresponding relationship between E3 and its substrates is challenging but required for understanding the regulatory network of ubiquitylation. The low abundance of ubiquitinated conjugates and high redu...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.9b00673
更新日期：2020-01-03 00:00:00
abstract：:Porcine reproductive and respiratory syndrome (PRRS) has devastated the pig industry worldwide for almost 25 years, and its virus (PRRSV) preferentially infects and replicates in pulmonary alveolar macrophages (PAMs). To discover cellular protein responses in PRRSV-infected PAMs, two-dimensional liquid chromatography-...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr201266z
更新日期：2012-05-04 00:00:00
abstract：:The final goal of the Russian part of the Chromosome-centric Human Proteome Project (C-HPP) was established as the analysis of the chromosome 18 (Chr 18) protein complement in plasma, liver tissue and HepG2 cells with the sensitivity of 10(-18) M. Using SRM, we have recently targeted 277 Chr 18 proteins in plasma, liv...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr300821n
更新日期：2013-01-04 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：:12-O-Tetradecanoyl-phorbol-13-acetate (TPA) is a plant derivative with multiple function as tumor promoter, differentiation revulsant or leukemia therapy drug. The molecular mechanism of its function is perplexing. Many studies have focused on the mechanism of TPA stimulation in tumor promotion of mouse models or term...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr0497677
更新日期：2005-03-01 00:00:00
abstract：:Recent physiological studies indicated that S. lividans metabolism was mainly glycolytic, whereas S. coelicolor metabolism was mainly oxidative. To determine whether such metabolic characteristics were correlated with consistent proteomics features, a comparative label-free, shotgun proteomics analysis of these strain...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.7b00163
更新日期：2017-07-07 00:00:00
abstract：:An important mechanism by which plants defend themselves against pathogens is the rapid execution of a hypersensitive response (HR). Tomato plants containing the Cf-4 resistance gene mount an HR that relies on the activation of phosphorylation cascades, when challenged with the Avr4 elicitor secreted by the pathogenic...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800619h
更新日期：2009-03-01 00:00:00
abstract：:Tomato fruit and seed allergens were detected by IgE-immunoblotting using sera from 18 adult tomato-sensitized patients selected based on a positive history skin prick test (SPT) and specific Immunglobulin (Ig) E-levels. Isolated tomato seed total protein showed high SPT activity comparable or even higher than tomato ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr800186d
更新日期：2009-03-01 00:00:00
abstract：:We report here using a novel technology-electrospray-assisted laser desorption ionization (ELDI)/mass spectrometry-for the rapid and sensitive detection of the major proteins that exist in dried biological fluids (e.g., blood, tears, saliva, serum), bacterial cultures, and tissues (e.g., porcine liver and heart) under...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr050442f
更新日期：2006-05-01 00:00:00
abstract：:Transient cerebral ischemia dramatically activates small ubiquitin-like modifier (SUMO2/3) conjugation. In cells exposed to 6 h of transient oxygen/glucose deprivation (OGD), a model of ischemia, SUMOylation increases profoundly between 0 and 30 min following re-oxygenation. To elucidate the effect of transient OGD on...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr200834f
更新日期：2012-02-03 00:00:00
abstract：:Pathogenesis of Staphylococcus aureus, an opportunistic human pathogen, is complex and involves many virulence factors including an array of surface proteins (adhesins) that promote bacterial interactions with extracellular matrix components. A better understanding of these interactions can be achieved by studying the...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr049866k
更新日期：2005-03-01 00:00:00
abstract：:Aberrant protein phosphorylation plays important roles in cancer-related cell signaling. With the goal of achieving multiplexed, comprehensive, and fully automated relative quantitation of site-specific phosphorylation, we present a simple label-free strategy combining an automated pH/acid-controlled IMAC procedure an...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr100394u
更新日期：2010-11-05 00:00:00
abstract：:Transferrin purification from mice serum samples by immunoaffinity chromatography (IAC) was optimized in order to study the possible modifications occurring in its glycans in collagen-induced arthritis (CIA) samples. SDS-PAGE and nanoLC-MS/MS were used to monitor the IAC purification performance. Afterward, a relative...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.0c00016
更新日期：2020-04-03 00:00:00
abstract：:Bone proteomics is an expanding field for understanding protein changes associated with disease as well as characterizing and detecting proteins preserved in fossil bone. Most previous studies have utilized a protocol with demineralization and extraction approach to isolate and characterize proteins from bone. Through...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.7b00670
更新日期：2018-01-05 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：:Direct tissue profiling and imaging mass spectrometry (MS) provides a detailed assessment of the complex protein pattern within a tissue sample. MALDI MS analysis of thin tissue sections results in over of 500 individual protein signals in the mass range of 2 to 70 kDa that directly correlate with protein composition ...
journal_title：Journal of proteome research
pub_type： 杂志文章,评审
doi：10.1021/pr0341282
更新日期：2004-03-01 00:00:00
abstract：:Bacillus Amyloliquefaciens SQR9 is a well-investigated plant growth-promoting rhizobacteria with strong root colonization capability. To identify the key proteins involved in in situ root colonization and biofilm formation, the proteomic profiles of planktonic and root colonized SQR9 cells were compared. A total of 75...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr500565m
更新日期：2014-12-05 00:00:00
abstract：:Sequence data from biomolecules such as DNA and proteins, which provide critical information for evolutionary studies, have been assumed to be forever outside the reach of dinosaur paleontology. Proteins, which are predicted to have greater longevity than DNA, have been recovered from two nonavian dinosaurs, but these...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00873
更新日期：2017-02-03 00:00:00
abstract：:The Mycobacterium tuberculosis membrane is rich in antigens that are potential targets for diagnostics and the development of new vaccines. To better understand the mechanisms underlying MTB virulence and identify new targets for therapeutic intervention, we investigated the differential composition of membrane proteo...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr400334k
更新日期：2013-12-06 00:00:00
abstract：:The development of medical interventions for the preservation of disease-free longevity would be facilitated by markers that predict healthy aging. Altered protein N-glycosylation patterns have been found with increasing age and several disease states. Here we investigate whether glycans derived from the total glycopr...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr1009959
更新日期：2011-04-01 00:00:00
abstract：:Since 2012, missing proteins (MPs) investigation has been one of the critical missions of Chromosome-Centric Human Proteome Project (C-HPP) through various biochemical strategies. On the basis of our previous testis MPs study, faster scanning and higher resolution mass-spectrometry-based proteomics might be conducive ...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/acs.jproteome.6b00390
更新日期：2016-11-04 00:00:00
abstract：:Advances in proteomics are continually driven by the introduction of new mass spectrometric instrumentation with improved performances. The recently introduced quadrupole Orbitrap (Q Exactive) tandem mass spectrometer allows fast acquisition of high-resolution higher-energy collisional dissociation (HCD) tandem mass s...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr3000249
更新日期：2012-06-01 00:00:00
abstract：:Bacillus weihenstephanensis is a subspecies of the Bacillus cereus sensu lato group of spore-forming bacteria known to cause food spoilage or food poisoning. The key distinguishing phenotype of B. weihenstephanensis is its ability to grow below 7 °C or, from a food safety perspective, to grow and potentially produce t...
journal_title：Journal of proteome research
pub_type： 杂志文章
doi：10.1021/pr501307t
更新日期：2015-05-01 00:00:00
