Name: sampling human indigenous saliva peptidome using a lollipop-like ultrafiltration probe: simplify and enhance peptide detection for clinical mass spectrometry
Uploaded: 2012-08-07T13:00:00
Description: Watch this Scientific Journal Video about Sampling Human Indigenous Saliva Peptidome Using a Lollipop-Like Ultrafiltration Probe: Simplify and Enhance Peptide Detection for Clinical Mass Spectrometry

यह काम राष्ट्रीय संस्थानों स्वास्थ्य अनुदान (R01-01-AI067395, R21-R022754-01, और R21 I58002-01) द्वारा समर्थित किया गया. हम सी. Niemeyer पांडुलिपि के महत्वपूर्ण पढ़ने के लिए धन्यवाद.

1. LLUF जांच का निर्माण <ol><li> polyethersulfone झिल्ली (2 2 सेमी) त्रिकोण polypropylene के paddles के साथ (कैलिफोर्निया विश्वविद्यालय, सैन डिएगो) paddles के सीमाओं पर epoxy के साथ gluing झिल्ली द्वारा सील किया गया. 30 केडीए में एक आणविक वजन कट ऑफ (MWCO...

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	<li>Denny, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+proteomes+of+human+parotid+and+ubmandibular/sublingual+gland+salivas+collected+as+the+ductal+secretions.">The proteomes of human parotid and ubmandibular/sublingual gland salivas collected as the ductal secretions.</a> J. Proteome Res. 7, 1994-2006 (2008).</li><li>Hu, S., Loo, J. A., Wong, D. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+saliva+proteome+analysis.">Human saliva proteome analysis.</a> Ann. N.Y. Acad. Sci. 1098, 323-329 (2007).</li><li>Ng, D. P., Koh, D., Choo, S. G., Ng, V., Fu, Q. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+storage+conditions+on+the+extraction+of+PCR-quality+genomic+DNA+from+saliva.">Effect of storage conditions on the extraction of PCR-quality genomic DNA from saliva.</a> Clin. Chim. Acta. 343, 191-194 (2004).</li><li>Wade, S. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+oral-diffusion-sink+device+for+extended+sampling+of+multiple+steroid+hormones+from+saliva.">An oral-diffusion-sink device for extended sampling of multiple steroid hormones from saliva.</a> Clin. Chem. 38, 1878-1882 (1992).</li><li>Hu, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large-scale+identification+of+proteins+in+human+salivary+proteome+by+liquid+chromatography/mass+spectrometry+and+two-dimensional+gel+electrophoresis-mass+spectrometry.">Large-scale identification of proteins in human salivary proteome by liquid chromatography/mass spectrometry and two-dimensional gel electrophoresis-mass spectrometry.</a> Proteomics. 5, 1714-1728 (2005).</li><li>Huang, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Comparative+proteomic+analysis+of+human+whole+saliva.">Comparative proteomic analysis of human whole saliva.</a> Arch. Oral Biol. 49, 951-962 (2004).</li><li>Guerrier, L., Lomas, L., Boschetti, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+simplified+monobuffer+multidimensional+chromatography+for+high-throughput+proteome+fractionation.">A simplified monobuffer multidimensional chromatography for high-throughput proteome fractionation.</a> J Chromatogr. A. 1073, 25-33 (2005).</li><li>Huang, C. M., Wang, C. C., Kawai, M., Barnes, S., Elmets, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Surfactant+sodium+lauryl+sulfate+enhances+skin+vaccination:+molecular+characterization+via+a+novel+technique+using+ultrafiltration+capillaries+and+mass+spectrometric+proteomics.">Surfactant sodium lauryl sulfate enhances skin vaccination: molecular characterization via a novel technique using ultrafiltration capillaries and mass spectrometric proteomics.</a> Mol. Cell Proteomics. 5, 523-532 (2006).</li><li>Huang, C. M., Wang, C. C., Kawai, M., Barnes, S., Elmets, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+protein+sampling+using+capillary+ultrafiltration+semi-permeable+hollow+fiber+and+protein+identification+via+mass+spectrometry-based+proteomics.">In vivo protein sampling using capillary ultrafiltration semi-permeable hollow fiber and protein identification via mass spectrometry-based proteomics.</a> J. Chromatogr. A. 1109, 144-151 (2006).</li><li>Huang, C. M., Wang, C. C., Barnes, S., Elmets, C. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+detection+of+secreted+proteins+from+wounded+skin+using+capillary+ultrafiltration+probes+and+mass+spectrometric+proteomics.">In vivo detection of secreted proteins from wounded skin using capillary ultrafiltration probes and mass spectrometric proteomics.</a> Proteomics. 6, 5805-5814 (2006).</li><li>Huang, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mass+spectrometric+proteomics+profiles+of+in+vivo+tumor+secretomes:+capillary+ultrafiltration+sampling+of+regressive+tumor+masses.">Mass spectrometric proteomics profiles of in vivo tumor secretomes: capillary ultrafiltration sampling of regressive tumor masses.</a> Proteomics. 6, 6107-6116 (2006).</li><li>Ahmed, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+approach+to+remove+albumin+for+the+proteomic+analysis+of+low+abundance+biomarkers+in+human+serum.">An approach to remove albumin for the proteomic analysis of low abundance biomarkers in human serum.</a> Proteomics. 3, 1980-1987 (2006).</li><li>Michishige, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+saliva+collection+method+on+the+concentration+of+protein+components+in+saliva.">Effect of saliva collection method on the concentration of protein components in saliva.</a> J. Med. Invest. 53, 140-146 (2006).</li><li>Kruger, C., Breunig, U., Biskupek-Sigwart, J., Dorr, H. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Problems+with+salivary+17-hydroxyprogesterone+determinations+using+the+Salivette+device.">Problems with salivary 17-hydroxyprogesterone determinations using the Salivette device.</a> Eur. J. Clin. Chem. Clin. Biochem. 34, 926-929 (1996).</li><li>Luque-Garcia, J. L., Neubert, T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sample+preparation+for+serum/plasma+profiling+and+biomarker+identification+by+mass+spectrometry.">Sample preparation for serum/plasma profiling and biomarker identification by mass spectrometry.</a> J. Chromatogr. A. 1153, 259-276 (2007).</li><li>Ramstrom, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Depletion+of+high-abundant+proteins+in+body+fluids+prior+to+liquid+chromatography+fourier+transform+ion+cyclotron+resonance+mass+spectrometry.">Depletion of high-abundant proteins in body fluids prior to liquid chromatography fourier transform ion cyclotron resonance mass spectrometry.</a> J. Proteome. Res. 4, 410-416 (2005).</li><li>Messana, I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Characterization+of+the+human+salivary+basic+proline-rich+protein+complex+by+a+proteomic+approach.">Characterization of the human salivary basic proline-rich protein complex by a proteomic approach.</a> J. Proteome. Res. 3, 792-800 (2004).</li><li>Li, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Possible+release+of+an+ArgGlyArgProGln+pentapeptide+with+innate+immunity+properties+from+acidic+proline-rich+proteins+by+proteolytic+activity+in+commensal+streptococcus+and+actinomyces+species.">Possible release of an ArgGlyArgProGln pentapeptide with innate immunity properties from acidic proline-rich proteins by proteolytic activity in commensal streptococcus and actinomyces species.</a> Infect. Immun. 68, 5425-5429 (2000).</li><li>Davtyan, T. K., Manukyan, H. A., Mkrtchyan, N. R., Avetisyan, S. A., Galoyan, A. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypothalamic+proline-rich+polypeptide+is+a+regulator+of+oxidative+burst+in+human+neutrophils+and+monocytes.">Hypothalamic proline-rich polypeptide is a regulator of oxidative burst in human neutrophils and monocytes.</a> Neuroimmunomodulation. 12, 270-284 (2005).</li><li>Jonsson, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gln-Gly+cleavage:+correlation+between+collision-induced+dissociation+and+biological+degradation.">Gln-Gly cleavage: correlation between collision-induced dissociation and biological degradation.</a> J. Am. Soc. Mass Spectrom. 12, 337-342 (2001).</li><li>Gibbons, R. J., Hay, D. I., Schlesinger, D. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Delineation+of+a+segment+of+adsorbed+salivary+acidic+proline-rich+proteins+which+promotes+adhesion+of+Streptococcus+gordonii+to+apatitic+surfaces.">Delineation of a segment of adsorbed salivary acidic proline-rich proteins which promotes adhesion of Streptococcus gordonii to apatitic surfaces.</a> Infect Immun. 59, 2948-2954 (1991).</li><li>Li, T., Johansson, I., Hay, D. I., Stromberg, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Strains+of+Actinomyces+naeslundii+and+Actinomyces+viscosus+exhibit+structurally+variant+fimbrial+subunit+proteins+and+bind+to+different+peptide+motifs+in+salivary+proteins.">Strains of Actinomyces naeslundii and Actinomyces viscosus exhibit structurally variant fimbrial subunit proteins and bind to different peptide motifs in salivary proteins.</a> Infect Immun. 67, 2053-2059 (1999).</li><li>Hardt, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Toward+defining+the+human+parotid+gland+salivary+proteome+and+peptidome:+identification+and+characterization+using+2D+SDS-PAGE,+ultrafiltration,+HPLC,+and+mass+spectrometry.">Toward defining the human parotid gland salivary proteome and peptidome: identification and characterization using 2D SDS-PAGE, ultrafiltration, HPLC, and mass spectrometry.</a> Biochemistry. 44, 2885-2899 (2005).</li><li>Wilmarth, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-dimensional+liquid+chromatography+study+of+the+human+whole+saliva+proteome.">Two-dimensional liquid chromatography study of the human whole saliva proteome.</a> J. Proteome Res. 3, 1017-1023 (2004).</li><li>Saitoh, E., Isemura, S., Sanada, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Complete+amino+acid+sequence+of+a+basic+proline-rich+peptide,+P-D,+from+human+parotid+saliva.">Complete amino acid sequence of a basic proline-rich peptide, P-D, from human parotid saliva.</a> J. Biochem. 93, 495-502 (1983).</li><li>Slomiany, B. L., Piotrowski, J., Czajkowski, A., Shovlin, F. E., Slomiany, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Differential+expression+of+salivary+mucin+bacterial+aggregating+activity+with+caries+status.">Differential expression of salivary mucin bacterial aggregating activity with caries status.</a> Int. J. Biochem. 25, 935-940 (1993).</li><li>Juarez, Z. E., Stinson, M. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+extracellular+protease+of+Streptococcus+gordonii+hydrolyzes+type+IV+collagen+and+collagen+analogues.">An extracellular protease of Streptococcus gordonii hydrolyzes type IV collagen and collagen analogues.</a> Infect Immun. 67, 271-278 (1999).</li><li>Lo, C. S., Hughes, C. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+characterization+of+a+protease+from+Streptococcus+oralis+C104.">Identification and characterization of a protease from Streptococcus oralis C104.</a> Oral Microbiol. Immunol. 11, 181-187 (1996).</li><li>Harrington, D. J., Russell, R. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+and+characterisation+of+two+extracellular+proteases+of+Streptococcus+mutans.">Identification and characterisation of two extracellular proteases of Streptococcus mutans.</a> FEMS Microbiol. Lett. 121, 237-241 (1994).</li><li>Huang, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+secretome+sampling+technology+for+proteomics.">In vivo secretome sampling technology for proteomics.</a> Proteomics Clin. Appl. 1, 953-962 (2007).</li><li>Skepo, M., Linse, P., Arnebrant, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Coarse-grained+modeling+of+proline+rich+protein+1+(PRP-1)+in+bulk+solution+and+adsorbed+to+a+negatively+charged+surface.">Coarse-grained modeling of proline rich protein 1 (PRP-1) in bulk solution and adsorbed to a negatively charged surface.</a> J. Phys. Chem. B. 110, 12141-12148 (2006).</li><li>Losic, D., Rosengarten, G., Mitchell, J. G., Voelcker, N. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pore+architecture+of+diatom+frustules:+potential+nanostructured+membranes+for+molecular+and+particle+separations.">Pore architecture of diatom frustules: potential nanostructured membranes for molecular and particle separations.</a> J. Nanosci. Nanotechnol. 6, 982-989 (2006).</li><li>Linhares, M. C., Kissinger, P. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacokinetic+monitoring+in+subcutaneous+tissue+using+in+vivo+capillary+ultrafiltration+probes.">Pharmacokinetic monitoring in subcutaneous tissue using in vivo capillary ultrafiltration probes.</a> Pharm Res. 10, 598-602 (1993).</li><li>Li, W., Hendrickson, C. L., Emmett, M. R., Marshall, A. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+intact+proteins+in+mixtures+by+alternated+capillary+liquid+chromatography+electrospray+ionization+and+LC+ESI+infrared+multiphoton+dissociation+Fourier+transform+ion+cyclotron+resonance+mass+spectrometry.">Identification of intact proteins in mixtures by alternated capillary liquid chromatography electrospray ionization and LC ESI infrared multiphoton dissociation Fourier transform ion cyclotron resonance mass spectrometry.</a> Anal. Chem. 71, 4397-4402 (1999).</li><li>Whitelegge, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protein-Sequence+Polymorphisms+and+Post-translational+Modifications+in+Proteins+from+Human+Saliva+using+Top-Down+Fourier-transform+Ion+Cyclotron+Resonance+Mass+Spectrometry.">Protein-Sequence Polymorphisms and Post-translational Modifications in Proteins from Human Saliva using Top-Down Fourier-transform Ion Cyclotron Resonance Mass Spectrometry.</a> Int. J. Mass Spectrom. 268, 190-197 (2007).</li><li>Castro, P., Tovar, J. A., Jaramillo, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adhesion+of+Streptococcus+mutans+to+salivary+proteins+in+caries-free+and+caries-susceptible+individuals.">Adhesion of Streptococcus mutans to salivary proteins in caries-free and caries-susceptible individuals.</a> Acta Odontol. Latinoam. 19, 59-66 (2006).</li><li>Challacombe, S. J., Sweet, S. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oral+mucosal+immunity+and+HIV+infection:+current+status.">Oral mucosal immunity and HIV infection: current status.</a> Oral Dis. 8, 55-62 (2002).</li><li>Jensen, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Salivary+acidic+proline-rich+proteins+in+rheumatoid+arthritis.">Salivary acidic proline-rich proteins in rheumatoid arthritis.</a> Ann. N.Y. Acad. Sci. 842, 209-211 (1998).</li></ol>

हमने पाया है कि कई पेप्टाइड टुकड़े पचाया नहीं मानव लार में मौजूद हैं. इन पेप्टाइड टुकड़ों प्रोलाइन युक्त प्रोटीन, actin, अल्फा amylase, एक ग्लोबिन अल्फा, बीटा ग्लोबिन, 1 histain, केरातिन 1, 7 mucin, polymeric इम्मुनोग्लोबुलिन रिस...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूची संख्या </td><td> टिप्पणियाँ </td></tr><tr><td> Polyethersulfone झिल्ली </td><td> पाल निगम </td><td></td><td> 30 केडीए MWCO </td></tr><tr><td> Teflon fluorinated के ethylene propylene ट्यूब </td><td> Upchurch वैज्ञानिक </td><td></td><td></td></tr><tr><td> ब्लू dextran </td><td> सिग्मा </td><td></td><td></td></tr><tr><td> नैनो नियंत्रण रेखा प्रणाली </td><td> Eksigent </td><td></td><td></td></tr><tr><td> C18 जाल स्तंभ </td><td> Agilent है </td><td> 5065-9913 </td><td></td></tr><tr><td> LTQ रैखिक जन आयन जाल स्पेक्ट्रोमीटर </td><td> थर्मो फिशर </td><td></td><td></td></tr><tr><td> 2 जादूगर </td><td> ऋषि - एन reseaआरसीएच </td><td></td><td></td></tr><tr><td> Acetonitrile - 0.1% चींटी एसिड </td><td> संयुक्त बेकर </td><td> 9832-03 </td><td> नियंत्रण रेखा / एमएस ग्रेड </td></tr><tr><td> जल 0.1% चींटी एसिड </td><td> संयुक्त बेकर </td><td> 9834-03 </td><td> नियंत्रण रेखा / एमएस ग्रेड </td></tr></tbody></table>

sampling human indigenous saliva peptidome using a lollipop-like ultrafiltration probe: simplify and enhance peptide detection for clinical mass spectrometry

Although human saliva proteome and peptidome have been revealed 1-2 they were majorly identified from tryptic digests of saliva proteins. Identification of indigenous peptidome of human saliva without prior digestion with exogenous enzymes becomes imperative, since native peptides in human saliva provide potential values for diagnosing disease, predicting disease progression, and monitoring therapeutic efficacy. Appropriate sampling is a critical step for enhancement of identification of human indigenous saliva peptidome. Traditional methods of sampling human saliva involving centrifugation to remove debris 3-4 may be too time-consuming to be applicable for clinical use. Furthermore, debris removal by centrifugation may be unable to clean most of the infected pathogens and remove the high abundance proteins that often hinder the identification of low abundance peptidome.
Conventional proteomic approaches that primarily utilize two-dimensional gel electrophoresis (2-DE) gels in conjugation with in-gel digestion are capable of identifying many saliva proteins 5-6. However, this approach is generally not sufficiently sensitive to detect low abundance peptides/proteins. Liquid chromatography-Mass spectrometry (LC-MS) based proteomics is an alternative that can identify proteins without prior 2-DE separation. Although this approach provides higher sensitivity, it generally needs prior sample pre-fractionation 7 and pre-digestion with trypsin, which makes it difficult for clinical use. 
To circumvent the hindrance in mass spectrometry due to sample preparation, we have developed a technique called capillary ultrafiltration (CUF) probes 8-11. Data from our laboratory demonstrated that the CUF probes are capable of capturing proteins in vivo from various microenvironments in animals in a dynamic and minimally invasive manner 8-11. No centrifugation is needed since a negative pressure is created by simply syringe withdrawing during sample collection. The CUF probes combined with LC-MS have successfully identified tryptic-digested proteins 8-11. In this study, we upgraded the ultrafiltration sampling technique by creating a lollipop-like ultrafiltration (LLUF) probe that can easily fit in the human oral cavity. The direct analysis by LC-MS without trypsin digestion showed that human saliva indigenously contains many peptide fragments derived from various proteins. Sampling saliva with LLUF probes avoided centrifugation but effectively removed many larger and high abundance proteins. Our mass spectrometric results illustrated that many low abundance peptides became detectable after filtering out larger proteins with LLUF probes. Detection of low abundance saliva peptides was independent of multiple-step sample separation with chromatography. For clinical application, the LLUF probes incorporated with LC-MS could potentially be used in the future to monitor disease progression from saliva.

Watch this Scientific Journal Video about Sampling Human Indigenous Saliva Peptidome Using a Lollipop-Like Ultrafiltration Probe: Simplify and Enhance Peptide Detection for Clinical Mass Spectrometry 

Sampling Human Indigenous Saliva Peptidome Using a Lollipop-Like Ultrafiltration Probe: Simplify and Enhance Peptide Detection for Clinical Mass Spectrometry

भविष्य नैदानिक ​​आवेदन के लिए लार के नमूने को ध्यान में रखते हुए, लॉलीपॉप की तरह एक ultrafiltration जांच (LLUF) के मानव मौखिक गुहा में फिट था गढ़े. NanoLC-LTQ मास स्पेक्ट्रोमेट्री से पचाया नहीं लार की प्रत्यक्ष विश्लेषण LLUF जांच की क्षमता के लिए बड़ी प्रोटीन और उच्च बहुतायत प्रोटीन हटाने के लिए, और कम प्रचुर पेप्टाइड्स अधिक से detectable का प्रदर्शन किया.

मानव स्वदेशी लार लॉलीपॉप की तरह एक ultrafiltration जांच का प्रयोग Peptidome नमूनाकरण: सरल और क्लीनिकल मास स्पेक्ट्रोमेट्री के लिए पेप्टाइड जांच बढ़ाएँ

Although human saliva proteome and peptidome have been revealed 1-2 they were majorly identified from tryptic digests of saliva proteins. Identification ...

Research

JoVE Journal

Medicine

Using Visual and Narrative Methods to Achieve Fair Process in Clinical Care

दृश्य और कथा तरीके का उपयोग करने के लिए क्लीनिकल देखभाल में उचित प्रक्रिया पाना

The Institute of Medicine has targeted patient-centeredness as an important area of quality improvement. A major dimension of patient-centeredness is ...

The &alpha;-test: Rapid Cell-free CD4 Enumeration Using Whole Saliva

The ÃƒÅ½Ã‚Â±-test: Rapid Cell-free CD4 Enumeration Using Whole Saliva

α परीक्षण: रैपिड सेल मुक्त सीडी 4 पूरे लार का प्रयोग गणन

There is an urgent need for affordable CD4 enumeration to monitor HIV disease. CD4 enumeration is out of reach in resource-limited regions due to the time ...

Collecting Saliva and Measuring Salivary Cortisol and Alpha-amylase in Frail Community Residing Older Adults via Family Caregivers

Salivary measures have emerged in bio-behavioral research  that are easy-to-collect, minimally invasive, and relatively inexpensive  biologic markers of ...

Quantitative Mass Spectrometric Profiling of Cancer-cell Proteomes Derived From Liquid and Solid Tumors

तरल और ठोस ट्यूमर से निकाली गई कैंसर सेल Proteomes के मात्रात्मक मास Spectrometric रूपरेखा

In-depth analyses of cancer cell proteomes are needed to elucidate oncogenic pathomechanisms, as well as to identify potential drug targets and diagnostic ...

Pre-clinical Orthotopic Murine Model of Human Prostate Cancer

पूर्व नैदानिक ​​Orthotopic Murine मानव प्रोस्टेट कैंसर के मॉडल

To study the multifaceted biology of prostate cancer, pre-clinical in vivo models offer a range of options to uncover critical biological information ...

A Detailed Protocol for Perspiration Monitoring Using a Novel, Small, Wireless Device

पसीना निगरानी के लिए एक विस्तृत प्रोटोकॉल एक उपन्यास का प्रयोग, छोटे, वायरलेस डिवाइस

Perspiration monitoring can be utilized for the detection of certain diseases, such as thermoregulation and mental disorders, particularly when the ...

Absolute Quantification of A&#946;1-42 in CSF Using a Mass Spectrometric Reference Measurement Procedure

Absolute Quantification of AÃƒÆ’Ã…Â½Ãƒâ€šÃ‚Â²1-42 in CSF Using a Mass Spectrometric Reference Measurement Procedure

Aβ के निरपेक्ष मात्रा<sub&gt; 1-42</sub&gt; सीएसएफ में प्रयोग एक मास Spectrometric संदर्भ माप प्रक्रिया

Alzheimer's disease (AD) is the most common neurodegenerative disease among the elderly and accounts for 60-80% of all cases of dementia. Currently, the ...

A Novel Clinical Grade Isolation Method for Human Kidney Perivascular Stromal Cells

मानव गुर्दा Perivascular Stromal कक्षों के लिए एक उपन्यास चिकित्सीय ग्रेड अलगाव विधि

Mesenchymal Stromal Cells (MSCs) are tissue homeostatic and immune modulatory cells that have shown beneficial effects in kidney diseases and ...

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

In Vitro and In Vivo Detection of Mitophagy in Human Cells, C. Elegans, and Mice

इन विट्रो में और Vivo में मानव कोशिकाओं में Mitophagy का पता लगाने, सी. एलिगेंस, और चूहों

Mitochondria are the powerhouses of cells and produce cellular energy in the form of ATP. Mitochondrial dysfunction contributes to biological aging and a ...

Surgical Training for the Implantation of Neocortical Microelectrode Arrays Using a Formaldehyde-fixed Human Cadaver Model

एक Formaldehyde-फिक्स्ड मानव शव मॉडल का उपयोग कर Neocortical Microelectrode arrays के आरोपण के लिए शल्य चिकित्सा प्रशिक्षण

This protocol describes a procedure to assist surgeons in training for the implantation of microelectrode arrays into the neocortex of the human brain. ...