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

この作品は、健康補助金の国立研究所（R01-AI067395-01、R21-R022754-01、およびR21-I58002-01）によってサポートされていました。私たちは、原稿の重要な読書のためにC.ニーマイヤーに感謝します。

1。 LLUFプローブの作成<ol><li>ポリエーテルスルホン膜（2 cm 2の ） はパドルの境界線上にエポキシ樹脂接着膜により三角形のポリプロピレン製パドル（カリフォルニア大学サンディエゴ校）で密封した。 30 kDaの分子量カットオフ（MWCO）と負に帯電したポリエーテルスルホン膜を用いた。 </li><li> LLUFプローブは20 mlの注射器に接続することができますので、テフロンフッ素化?...

<ol>
	<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>

我々は、多くのペプチドフラグメントは、人間の消化唾液中に存在することを発見した。これらのペプチド断片は、プロリンリッチタンパク質、アクチン、α-アミラーゼ、α1グロビン、βグロビン、histain 1、ケラチン1、ムチン7、高分子免疫グロブリン受容体、satherin、S100A9の様々なフォームからの誘導体である。未定の切断部位を有するペプチドの生産に貢献する多くの要因があるかもしれ?...

<table border="1"><tbody><tr><td> 試薬の名前 </td><td> 会社 </td><td> カタログ番号 </td><td> コメント </td></tr><tr><td>ポリエーテルスルホン膜</td><td>ポール·コーポレーション</td><td></td><td> 30kDaのMWCO </td></tr><tr><td>テフロンフッ素化エチレンプロピレン管</td><td>アップチャーチ科学</td><td></td><td></td></tr><tr><td>ブルーデキストラン</td><td>シグマ</td><td></td><td></td></tr><tr><td>ナノLCシステム</td><td> Eksigent </td><td></td><td></td></tr><tr><td> C18トラップカラム</td><td>アジレント</td><td> 5065-9913 </td><td></td></tr><tr><td> LTQリニアイオントラップ型質量分析計</td><td>サーモフィッシャー</td><td></td><td></td></tr><tr><td>魔術師2 </td><td> SAGE-N ReseaRchの</td><td></td><td></td></tr><tr><td>アセトニトリル0.1％ギ酸</td><td> JTベーカー</td><td> 9832から03 </td><td> LC / MSグレード</td></tr><tr><td>水0.1％ギ酸</td><td> JTベーカー</td><td> 9834から03 </td><td> LC / MSグレード</td></tr></tbody></table>

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

ヒト唾液プロテオームとペプチドーが1-2を明らかにされているが、それらはすぎなかったトリプシン唾液タンパク質の消化物から同定された。外因性の酵素による事前消化せずにヒト唾液の先住民族ペプチドーの同定は、ヒト唾液でネイティブペプチドが疾患の進行を予測し、治療効果を監視し、病気を診断するための潜在的な値を提供するので、必須となります。適切なサンプリングは、人間の土着の唾液ペプチドーの識別の強化のための重要なステップです。 3から4の残骸を削除するには、遠心分離を含むヒト唾液をサンプリングする従来の方法は時間がかかりすぎるの臨床使用のために適用可能であることかもしれません。さらに、遠心分離による瓦礫撤去は、感染した病原体のほとんどをきれいにし、多くの場合、低濃度のペプチドーの識別を妨げる高濃度タンパク質を除去することができない場合があります。 PRI、従来のプロテオミクス的手法marilyゲル内消化との結合の二次元ゲル電気泳動（2-DE）ゲルは多くの唾液タンパク質の5-6を識別することができる利用しています。しかし、このアプローチは、一般的に低濃度のペプチド/タンパク質を検出するのに十分な小文字は区別されません。液体クロマトグラフィー - 質量分析（LC-MS）ベースのプロテオミクス前の2-DE分離することなくタンパク質を識別することができる代替手段です。このアプローチは、より高い感度を提供していますが、それは一般的に臨床使用が困難になってトリプシンと以前のサンプルプレ分画7とプレ消化を必要とします。 サンプル調製のために質量分析の妨害を回避するために、我々は、キャピラリ限外濾過（CUF）プローブ8から11と呼ばれる技術を開発しました。我々の研究室からのデータは、CUFプローブは、ダイナミックかつ低侵襲な方法8の動物の様々な微小環境から、生体内でタンパク質を捕捉することが可能であることを実証-11。負圧は、サンプル収集中に撤退単に注射器によって作成されているので全く遠心分離は必要ありません。 LC-MSを組み合わせたCUFプローブが成功したトリプシン消化し ​​たタンパク質の8-11を同定した。本研究では、簡単に人間の口腔内に収まることができロリポップのような限外濾過（LLUF）プローブを作成することにより、限外ろ過サンプリング技術をアップグレードしました。トリプシン消化せずにLC-MSによる直接分析では、国産ヒト唾液は様々なタンパク質に由来する多くのペプチド断片が含まれていることを示した。 LLUFプローブと唾液をサンプリングする遠心分離を避けますが、効果的に多くの大規模、高濃度タンパク質を除去。私たちの質量分析の結果は、多くの低濃度のペプチドはLLUFプローブで大きなタンパク質をフィルタリングした後に検出されなったことを示す。低濃度の唾液ペプチドの検出は、クロマトグラフィーを持つ複数のステップのサンプルの分離独立していた。臨床応用のために、LLUFプローブが組み込まれてLC-MSとdが潜在的に唾液から疾患の進行を監視するために、将来的に使用することができます。

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

将来の臨床応用のための唾液採取を考慮して、ロリポップのような限外濾過（LLUF）プローブは、ヒトの口腔内に収まるように作製した。ナノLC-LTQ質量分析法による消化唾液の直接分析は、大規模なタンパク質や高濃度タンパク質を除去し、低豊富なペプチドは多くの検出可能なようにするLLUFプローブの能力を実証した。

ロリポップライク限外プローブを用いたヒト先住民唾液ペプチドーをサンプリング：臨床質量分析のためのペプチドの検出を簡素化し、強化

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

臨床ケアのフ​​ェアプロセスを達成するために、Visualと物語メソッドの使用

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-テスト：全唾液を用いた迅速な無細胞CD4列挙

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

唾液を収集し、家族介護者を経由して高齢者を居住する虚弱コミュニティ唾液コルチゾールおよびα-アミラーゼの測定

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

液体および固形腫瘍由来の癌細胞のプロテオームの定量的質量分析プロファイリング

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

ヒト前立腺癌の前臨床同所性マウスモデル

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

汗は小説、小型、ワイヤレスデバイスを使用した監視するための詳細なプロトコル

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の絶対定量<sub&gt; 1-42</sub&gt; CSFに質量分析リファレンス測定手順を使用して

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

ひと腎臓血管周囲間質細胞の新規臨床等級分離法

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

In VitroとIn Vivo検出、ひと細胞、線虫、マウス Mitophagy の

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

人間の死体をホルマリン固定モデルを用いた皮質微小電極アレイの注入の手術トレーニング

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