Abstract:
:In this paper, we describe a method for analyzing large-volume protein samples using capillary electrophoresis in conjunction with laser-induced fluorescence detection (CE-LIF). To improve the stacking and separation efficiencies of proteins, we added either 0.01% sodium dodecyl sulfate (SDS) or 0.01% poly(ethylene oxide) (PEO) to the Tris-borate solutions (pH 10.0) used to prepare the protein samples. After injection of the large-volume samples (ca. 1.0 microL, 0.1 microM), the proteins migrate against the electroosmotic flow (EOF) and enter the PEO zone; this process causes them to slow and stack at the boundary between the PEO and sample zones. As a result, the limits of detection (LODs) at a signal-to-noise (S/N) of 3 for most proteins are sub-nM to several nM. For instance, the LOD (S/N = 3) for alpha-lactalbumin is 0.48 nM, which is an 84-fold sensitivity enhancement over the traditional method. By applying a short plug of 0.2% SDS prior to sample injection, a greater number of peaks, representing the microheterogeneity of the proteins, were resolved and the stacking efficiency of the proteins increased slightly. This method allowed us to detect 12 peaks when injecting a large volume of sample containing six model proteins (0.1 microM). We also analyzed the microheterogeneities of the proteins by using CE with UV-Vis absorption detection when injecting a large volume of sample containing six model proteins (1.0 microM) in the presence of a 1.0% SDS plug. The practical method is validated by the detection of human serum albumin in a urine sample, obtained from a healthy female, without sample pretreatment; its concentration was 0.18 microM. We further demonstrate the capability of this method to detect low amounts of proteins through the detection of 45 nM hemoglobin after injecting ca. 1.0 microL of ultradilute lysed red blood cells. The experimental results indicate that our proposed method has great potential for use in diagnosis and proteomics applications.
journal_name
J Proteome Resjournal_title
Journal of proteome researchauthors
Huang YF,Hsieh MM,Tseng WL,Chang HTdoi
10.1021/pr050298ykeywords:
subject
Has Abstractpub_date
2006-02-01 00:00:00pages
429-36issue
2eissn
1535-3893issn
1535-3907journal_volume
5pub_type
杂志文章abstract::Colon cancer is the most common cancer and the third leading cause of cancer mortality in humans. Using mass spectrometry-based metabolomics, the current study revealed the accumulation of four uremic toxins (cresol sulfate, cresol glucuronide, indoxyl sulfate, and phenyl sulfate) in the serum of mice harboring adenom...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr5013152
更新日期:2015-04-03 00:00:00
abstract::The localization of proteins at a tissue- or cell-type-specific level is tightly linked to the protein function. To better understand each protein's role in cellular systems, spatial information constitutes an important complement to quantitative data. The standard methods for determining the spatial distribution of p...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.0c00486
更新日期:2020-12-04 00:00:00
abstract::The fungal pathogen Fusarium graminearum (telomorph Gibberella zeae) is the causal agent of several destructive crop diseases. Identifying interactions among F. graminearum proteins and understanding their functions can provide insights into pathogenic mechanisms underlying F. graminearum-host interactions. F. gramine...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr900415b
更新日期:2009-10-01 00:00:00
abstract::Isobaric tagging is the method of choice in mass-spectrometry-based proteomics for comparing several conditions at a time. Despite its multiplexing capabilities, some drawbacks appear when multiple experiments are merged for comparison in large sample-size studies due to the presence of missing values, which result fr...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.8b00110
更新日期:2018-06-01 00:00:00
abstract::Adipose tissue is a major storage site for lipophilic environmental contaminants. The environmental metabolic disruptor hypothesis postulates that some pollutants can promote obesity or metabolic disorders by activating nuclear receptors involved in the control of energetic homeostasis. In this context, monoethylhexyl...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200765v
更新日期:2011-12-02 00:00:00
abstract::Proteoforms containing post-translational modifications (PTMs) represent a degree of functional diversity only harnessed through analytically precise simultaneous quantification of multiple PTMs. Here we present a method to accurately differentiate an unmodified peptide from its PTM-containing counterpart through data...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.0c00685
更新日期:2020-10-02 00:00:00
abstract::Candida albicans (C. albicans) is a worldwide cause of fungal infectious diseases. As a general post-translational modification (PTM), lysine acetylation of proteins play an important regulatory role in almost every cell. In our research, we used a high-resolution proteomic technique (LC-MS/MS) to present the comprehe...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00052
更新日期:2016-08-05 00:00:00
abstract::The International Mouse Phenotyping Consortium program has been established to ascribe biological functions to systematically knocked-out (KO) genes by in vivo and ex vivo phenotyping. The plasma clinical chemistry screen includes an assessment of liver, kidney, and bone function and provides a basic lipid profile and...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr501039k
更新日期:2015-05-01 00:00:00
abstract::Francisella tularensis is a Gram-negative bacterium that causes the zoonotic disease tularemia. The historical development of tularemia as a biological weapon has led to it being characterized by the CDC as a category A biothreat agent. Neither posttranslational modification (PTM) of proteins, in particular lysine ace...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.9b00512
更新日期:2020-04-03 00:00:00
abstract::Ion mobility-mass spectrometry (IM-MS) is a useful technique for determining information about analyte ion conformation in addition to mass/charge ratio. The physical principles that govern the mobility of an ion through a gas in the presence of a uniform electric field are well understood, enabling rotationally avera...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr3005327
更新日期:2012-11-02 00:00:00
abstract::The endothelium plays a critical role in orchestrating the inflammatory response seen during sepsis. Many of the inflammatory effects of Gram-negative sepsis are elicited by lipopolysaccharide (LPS), a glycolipid component of bacterial cell walls. Lipid-rich microdomains have been shown to concentrate components of th...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr049824w
更新日期:2005-03-01 00:00:00
abstract::Epithelial ovarian carcinoma has in general a poor prognosis since the vast majority of tumors are genomically unstable and clinically highly aggressive. This results in rapid progression of malignancy potential while still asymptomatic and thus in late diagnosis. It is therefore of critical importance to develop meth...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr201258q
更新日期:2012-05-04 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::To assess the global effect of preterm birth on fetal metabolism and maternal-fetal nutrient transfer, we used a mass spectrometric-based chemical phenotyping approach on cord blood obtained at the time of birth. We sampled umbilical venous, umbilical arterial, and maternal blood from mothers delivering very-low birth...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr400122v
更新日期:2013-06-07 00:00:00
abstract::The study of glycosylation patterns (glycomics) in biological samples is an emerging field that can provide key insights into cell development and pathology. A current challenge in the field of glycomics is to determine how to quantify changes in glycan expression between different cells, tissues, or biological fluids...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr070476i
更新日期:2008-01-01 00:00:00
abstract::Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and multiple reaction monitoring mass spectrometry (MRM-MS) proteomics analyses were performed on eccrine sweat of healthy controls, and the results were compared with those from individuals diagnosed with schizophrenia (SZ). This is the first large scale study...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr2007957
更新日期:2012-04-06 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::Deficiency of the sulfide metabolizing protein ETHE1 is the cause of ethylmalonic encephalopathy (EE), an inherited and severe metabolic disorder. To study the molecular effects of EE, we performed a proteomics study on mitochondria from cultured patient fibroblast cells. Samples from six patients were analyzed and re...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr101218d
更新日期:2011-05-06 00:00:00
abstract::Equine grass sickness (EGS) is a frequently fatal disease of horses, responsible for the death of 1 to 2% of the U.K. horse population annually. The etiology of this disease is currently uncharacterized, although there is evidence it is associated with Clostridium botulinum neurotoxin in the gut. Prevention is current...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.7b00784
更新日期:2018-03-02 00:00:00
abstract::By foliar fortification with selenite, selenium (Se)-enriched rice with a higher Se content and grain yield has been generated. However, the regulatory mechanisms of Se response in rice grains remain unknown; therefore, we carried out a comparative proteomics study in Se-enriched rice grains by using two approaches in...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr300878y
更新日期:2013-02-01 00:00:00
abstract::A proteoform family is a group of related molecular forms of a protein (proteoforms) derived from the same gene. We have previously described a strategy to identify proteoforms and elucidate proteoform families in complex mixtures of intact proteins. The strategy is based upon measurements of two properties for each p...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.7b00516
更新日期:2017-11-03 00:00:00
abstract::Human saliva is a protein-rich, easily accessible source of potential local and systemic biomarkers to monitor changes that occur under pathological conditions; however, little is known about the changes in abundance associated with normal aging. In this study, we performed a comprehensive proteomic profiling of poole...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr900478h
更新日期:2009-11-01 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::An efficient means for the identification of prognostic and predictive biomarkers is essential in today's cancer management. A new approach toward biomarker discovery has therefore been proposed, where pathways instead of individual proteins would be monitored and targeted. Recently, the 'secretome', a biological flui...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr8008572
更新日期:2009-03-01 00:00:00
abstract::The chromatophore membrane of the photosynthetic diazotroph Rhodospirillum rubrum is of vital importance for a number of central processes, including nitrogen fixation. Using a novel amphiphile, we have identified protein complexes present under different nitrogen availability conditions by the use of two-dimensional ...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr100838x
更新日期:2011-06-03 00:00:00
abstract::Akt1 is a serine/threonine kinase that promotes cell growth and survival. Previously, Akt1 activation in a double transgenic (DTG) mouse model fed a high-fat/high-sucrose (HF/HS) diet was found to promote type IIb muscle growth and to lead to a significant reduction in obesity. Here, we have used metabolomics to exami...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr500756u
更新日期:2015-01-02 00:00:00
abstract::We undertook an unbiased metabolite profiling of fibroblasts from schizophrenia patients and healthy controls to identify metabolites and pathways that are dysregulated in disease, seeking to gain new insights into the disease biology of schizophrenia and to discover potential disease-related biomarkers. We measured p...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.6b00628
更新日期:2017-02-03 00:00:00
abstract::Reference materials are vital to benchmarking the reproducibility of clinical tests and essential for monitoring laboratory performance for clinical proteomics. The reference material utilized for mass spectrometric analysis of the human proteome would ideally contain enough proteins to be suitably representative of t...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/acs.jproteome.8b00165
更新日期:2018-06-01 00:00:00
abstract::Kalirin-7 (Kal7), a multifunctional Rho GDP/GTP exchange factor (GEF) for Rac1 and RhoG, is embedded in the postsynaptic density at excitatory synapses, where it participates in the formation and maintenance of dendritic spines. Kal7 has been implicated in long-term potentiation, fear memories, and addiction-like beha...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr200088w
更新日期:2011-06-03 00:00:00
abstract::A novel linear ion trap (LIT) mass spectrometer with dual matrix-assisted laser desorption/ionization (MALDI) and electrospray ionization (ESI) ionization sources has been built in the MALDI-LIT-ESI configuration. The design features two independent ion source/ion optical channels connected to opposite ends of a singl...
journal_title:Journal of proteome research
pub_type: 杂志文章
doi:10.1021/pr060514i
更新日期:2007-02-01 00:00:00