專利名稱：：用于診斷和評價血管病的多肽標志物的制作方法用于診斷和評價血管病的多肽標志物本發(fā)明涉及來自受試者之樣品是否存在一種或多種肽標志物用于診斷和評價血管病(vasculardisease,VD)之嚴重程度的用途，以及涉及診斷和評價這些血管病的方法，其中是否存在所述肽標志物標志著VD的嚴重程度。血管病是侵襲有機體血管并因此侵襲器官比如心臟、腦、腎等的疾病。它們包括例如動脈硬化、循環(huán)障礙、高血壓和心律失常。血管動脈硬化指由血管沉積物引起動脈硬化。膽固醇結(jié)晶的沉積物導致炎性病灶(動脈粥樣化)的形成，血液成分、脂質(zhì)、代謝殘渣(metabolicslags)以及石灰鹽(limesalts)易于沉積在所述炎性病灶中。形成所謂的斑(二維硬化)，因此血管壁變得較硬以及較為狹窄。動脈失去其彈性并且難于執(zhí)行其任務(wù)(即將血液從心臟運輸?shù)缴眢w的各個區(qū)域)。繼發(fā)性疾病包括例如心絞痛、心肌梗塞、循環(huán)性虛脫、中風。循環(huán)障礙主要影響身體下部，從腹主動脈到足動脈，導致減少血流和氧氣向肌肉組織的供應(yīng)，所述肌肉組織逐漸變?yōu)閴乃?。在晚期，潰瘍形成并阻塞血管至截肢變得不可避免的程度。高血壓具有不確定性病因，因此，攝取藥物或過量分泌腎上腺激素可導致血壓增高。高血壓還可出現(xiàn)在持續(xù)性緊張(permanentsress)中，其導致血管痙攣。高血壓損害血管壁，因此具有破裂或阻塞的風險。如果心跳的規(guī)律性被擾亂，該病癥被稱為心律失常。心跳可能太快(心動過速(tachycardia))、太慢(心搏緩慢(bradycardia))或不規(guī)律(心率失常)。血管病可通過預防而避免，因為其也是由不健康或不正常的生活行為引起的。通過根本性轉(zhuǎn)變生活方式，動脈硬化可在早期階段被阻止，例如通過降低血壓和血脂水平。血管病的進程可另外通過藥物治療而變慢(例如乙酰水楊酸、P受體阻斷劑、ACE抑制劑等)。然而，應(yīng)當注意，被損傷的血管是不可修復的，并且晚期的進程是不可逆的。因此，血管病的早期檢測尤其重要。心臟在冠心病中，首先間接通過評價風險因子以及通過非侵入式檢查實現(xiàn)VD的診斷，所述非侵入式檢查比如血壓測量、靜息心電圖和運動心電圖、以及以檢測下列項的血像脂質(zhì)狀態(tài)(LDL膽固醇、HDL膽固醇、甘油三酸酯)、空腹血液葡萄糖水平和(如果需要)HbAlc。如果這樣的檢查得出存在高風險特征，即預期在不遠的將來發(fā)生嚴重的血管事件(死亡、心肌梗塞)，那么利用非侵入式診斷進行更準確的診斷，例如以導管檢查或冠狀動脈造影術(shù)的形式。因此，利用導管或X-射線法檢查心臟和冠狀血管以及其它血管。X-射線造影劑用于在X-射線圖像上更好地顯現(xiàn)心臟和血管。冠狀動脈造影術(shù)的適用者包括初步測試可能性(preliminarytestprobability)為低度或中度但非侵入式i^斷未能提供可靠結(jié)果、由于缺陷或疾病不可能進行非侵入式檢查的患者、以及由于工作相關(guān)原因必須排除可能存在冠心病的患者(例如飛行員、消防員)。然而，除了上述提及的初步檢查，只要各種并發(fā)癥比如甲狀腺功能亢進或?qū)υ煊皠┑淖儜B(tài)反應(yīng)被排除，即可實施冠狀動脈造影術(shù)。此外，由于造影劑通過腎臟分泌，必須保證足夠的腎功能，或者對于透析依賴性的患者，必須總是在檢查之后接著實施透析。因此，很明顯需要對早期的和可靠的診斷血管病的非侵入式可能性。腎臟腎臟的血管病包括腎動脈狹窄腎動脈血栓形成腎動脈栓塞腎靜脈血栓形成腎動脈狹窄是腎動脈或其主要分支的單側(cè)或雙側(cè)狹窄。它可能是動脈高血壓的原因，因而所述動脈高血壓被稱為腎血管性高血壓。在約70%的病例中，它的原因是動脈硬化(在高齡中是最主要的)，以及在約20。/。的病例中，其原因是肌纖維結(jié)構(gòu)不良(結(jié)締組織異常)。少見地，其原因包括主動脈或腎動脈的動脈瘤、血管炎、腫瘤或嚢肺的機械壓迫、栓塞或血栓形成。腎動脈狹窄導致通過受影響腎臟的血流減少。為了彌補推測的(局部！)血壓降低，腎臟增加腎素的產(chǎn)生，腎素通過血管緊張素-醛固酮機制導致整個有機體的血液體積增加和血壓升高，并且導致動脈高血壓。因此，當高血壓逐漸形成時，通常發(fā)現(xiàn)腎動脈狹窄，但是所有高血壓中僅僅約1~2%由此引起。對于治療，具有不同的可能性PTA(經(jīng)皮穿刺導管血管形成術(shù)(Percutaneoustransluminalcatheterangioplasty)):利用插入球?qū)Ч軘U張狹窄(球擴張)；支架插入絲網(wǎng)(支架)以保持血管開放；手術(shù)消除狹窄。腎動脈血栓形成的常見原因是源自心臟的血栓，例如，在心房纖維顫動期間，所述心房纖維顫動伴隨癥狀例如脅腹痛、蛋白尿、非常高的LDH。還在腎靜脈血栓形成中觀察到脅腹痛，但是除了蛋白尿以外，在一些情況下，還觀察到血尿或腎病綜合征。腦腦區(qū)域中狹窄的血管導致氧供應(yīng)減少，并且當動脈被阻塞時(例如被動脈硬化之變化導致的急性血塊所阻塞)，出現(xiàn)中風并伴隨知覺喪失、癱瘓、言語障礙等。在腦動脈中，比如在大動脈中，動脈硬化可在極少情況下導致血管壁動脈瘤，并且與風險因子例如高血壓一起，血管壁可能破裂并引起危及生命的內(nèi)出血。令人驚訝地，現(xiàn)在已經(jīng)發(fā)現(xiàn)來自受試者的尿樣品中的特殊肽標志物可用于VD的診斷并因此判定是否需要藥物治療。因此，本發(fā)明涉及利用來自受試者的尿樣品中至少一種肽標志物(理想地是幾種多肽標志物)的存在與否診斷血管病的用途，其中所述一種或多種多肽標志物選自多肽標志物至No.526,所述多肽標志物No.l至No.526的特征在于具有如表1所述的分子質(zhì)量和遷移時間。<table>tableseeoriginaldocumentpage13</column></row><table><table>tableseeoriginaldocumentpage14</column></row><table><table>tableseeoriginaldocumentpage15</column></row><table>優(yōu)選地，4吏用標志物1104和/或107~413。使用本發(fā)明，也可以測定VD的嚴重程度。此信息有助于決定使用怎樣的治療措施。利用毛細管電泳(CE)測定遷移時間，例如，如實施例笫2項中所述。因此，在30kV的電壓下操作長度為90cm、內(nèi)徑(ID)為75pm以及外徑(OD)為360pm的玻璃毛細管。使用含有30%甲醇、0.5%曱酸的水作為樣品的溶劑。已知CE遷移時間可發(fā)生變化。不過，對于所用的任何CE系統(tǒng)，多肽標志物洗脫的順序通常是相同的。為了平衡遷移時間的差異，可使用已知遷移時間的標準校正所述系統(tǒng)。這些標準可以是例如實施例中所述的多肽(見實施例，第3項)。利用毛細管電泳-質(zhì)i普(CE-MS)測定表1至3中所示多肽標志物的特征，所述毛細管電泳-質(zhì)譜(CE-MS)方法已經(jīng)詳細描述于，例如Neuhoff等(RapidCommunicationsinmassspectrometry,2004,Vol，20pp.l49-156)。各個測量之間或不同質(zhì)i普儀之間的分子質(zhì)量變化相對較小，典型地在±0.1%的范圍內(nèi)，優(yōu)選在±0.05%的范圍內(nèi)，更優(yōu)選在±0.03%的范圍內(nèi)，甚至更優(yōu)選在±0.01%的范圍內(nèi)。根據(jù)本發(fā)明的多肽標志物是蛋白質(zhì)或肽或者蛋白質(zhì)或肽的降解產(chǎn)物。它們可被化學修飾，例如通過翻譯后修飾，比如糖基化、磷酸化、烷基化或二硫鍵，或通過例如在降解范圍內(nèi)的其它反應(yīng)。此外，多肽標志物還可在樣品純化的范圍內(nèi)被化學改變，例如被氧化。從測定多肽標志物的參數(shù)(分子質(zhì)量和遷移時間)著手，可以通過現(xiàn)有4支術(shù)7>知的方法確定相應(yīng)多肽的序列。根據(jù)本發(fā)明的多肽(見表1至4)用于診斷VD的嚴重程度。"診斷，，指通過將癥狀或現(xiàn)象歸因于疾病或損傷而獲取知識的過程。在目前情況下，VD的嚴重程度由特殊多肽標志物的存在與否推出。因此，在來自受試者的樣品中測定根據(jù)本發(fā)明的多肽標志物，其中它的存在與否可用于推斷VD的嚴重程度。多肽標志物的存在與否可以通過現(xiàn)有技術(shù)的任何公知方法進行測量。下面舉例說明可能的公知方法。如果多肽標志物的測量值至少跟其閾值一樣高，那么認為存在該多肽標志物。如果測量值更低，那么^人為不存在多肽標志物?？赏ㄟ^測量方法的靈敏度(檢測限度)或者根據(jù)經(jīng)驗確定閾值。在本發(fā)明的上下文中，如果樣品中針對某一分子質(zhì)量的測量值至少是空白樣品(例如，僅僅緩沖劑或溶劑)的兩倍，優(yōu)選地認為超過閾值。以測定多肽標志物的存在與否的方式使用該多肽標志物，其中存在與否標志著VD的嚴重程度(頻率標志物(frequencymarker))。因此，有些多肽標志物通常存在于患有VD的受試者中，但是較少見或不存在于未患有VD的受試者中，例如，1-24(表2)。此外，有些多肽標志物存在于VD患者中，比如多肽標志物No.25~106，但是較少見地或根本不存在于未患有VD的患者中。表2:用于診斷血管病的多肽標志物(頻率標志物)、其分子質(zhì)量和遷移時間、以及以因子形式表示其是否存在于VD患者(VD)和對照組(對照)中(1=100%,0=0%;如實施例中所述樣品處理和測量)<table>tableseeoriginaldocumentpage17</column></row><table>頻率標志物(測定存在與否)之外或者作為其替代，如表3所述的幅度標志物也可用于VD的診斷(Nos.l07~526)。在使用幅度標志物的方式中，其存在與否并非至關(guān)重要，但是如果兩組中都存在信號，那么信號的高度(幅度)起決定作用。在表3和4中，描述了相對所有測量樣品進行平均的相應(yīng)信號(以質(zhì)量和遷移時間為特征)的平均幅度。兩種歸一化方法可以實現(xiàn)不同濃縮樣品之間或不同測量方法之間的可比性。在第一種方法中，樣品的所有肽信號被歸一化為l百萬計數(shù)的總幅度。因此，各個標志物的各自平均幅度被規(guī)定為份數(shù)/百萬(ppm)。通過該方法得到的幅度標志物被顯示在表3中(Nos.107413)。此外，可通過作為替代的歸一化方法定義另外的幅度標志物在這種情況下，利用共用歸一化因子衡量一個樣品的所有肽信號。因此，單個樣品的肽幅度和所有已知多肽的參照值之間形成線性回歸?；貧w線的斜率正好對應(yīng)于相對濃度并且被用作該樣品的歸一化因子。通過該歸一化方法得到的生物標志物凈皮顯示在表4中(Nos.414~526)。所有所用組由至少20個不同患者的樣品或?qū)φ諛悠方M成以得到可靠的平均幅度。以下述函數(shù)作出診斷決定(是否為VD):與對照組或VD組的平均幅度相比，患者樣品中各自多肽標志物的幅度是多高。如果幅度更相當于VD組的平均幅度，認為存在血管病，以及如果幅度更相當于對照組的平均幅度，認為不存在VD。測量值和平均幅度之間的距離可被認為是樣品屬于某一組的概率。將利用標志物No.137給出示例性解釋(表3)。VD中所述標志物的平均幅度顯著增加(12044ppm相對于對照組中的5726ppm)?，F(xiàn)在，如果患者樣品中該標志物的值為0至5726ppm或最多超出該范圍20%,即從0至6871ppm，那么該樣品屬于對照組。如果值是12044ppm或高出低于其20。/o的值，或更高，即在9635和非常高的值之間，則凈皮認為是血管病的指征。作為替代，測量值和平均幅度之間的距離可被認為是樣品屬于某一組的概率。在一些樣品中，頻率標志物是低幅度的幅度標志物的變體。通過將其中沒有發(fā)現(xiàn)標志物的相應(yīng)樣品包括到非常小之幅度(檢測限度級別)的幅度計算中，可能將這樣的頻率標志物轉(zhuǎn)化成幅度標志物。表3:才艮據(jù)方法1歸一化的幅度標志物<table>tableseeoriginaldocumentpage19</column></row><table><table>tableseeoriginaldocumentpage20</column></row><table>表4:根據(jù)方法2歸一化的幅度標志物<table>tableseeoriginaldocumentpage21</column></row><table>已測定一種或多種多肽標志物存在與否的樣品的來源受試者可以是可能患VD的任何受試者。優(yōu)選地，所述受試者是哺乳動物，并且最優(yōu)選地，是人。在本發(fā)明的一個優(yōu)選實施方案中，不只一種多肽標志物，而是多肽標志物的組合用于測定VD的嚴重程度，其中VD的嚴重程度可從所述多肽標志物的存在與否進行推斷。通過比較多種多肽標志物，可降低或避免由于患者或?qū)φ諅€體中通常存在可能性的一些個體偏差導致的總體結(jié)果中的偏差。所述在其中測量根據(jù)本發(fā)明的肽標志物存在與否的樣品可以是從受試者身體中得到的任何樣品。所述樣品是含有適于提供有關(guān)受試者狀態(tài)(是否是VD)之信息的多肽組合物的樣品。例如，其可以是血液、尿、滑液、組織液、身體分泌物(bodysecretion)、汗、腦脊液、淋巴、腸液、胃液或胰液、膽汁、淚液、組織樣品、精液、陰道液或糞便樣品。優(yōu)選地，其是液體樣品。在一個優(yōu)選實施方案中，所述樣品是尿樣品或血液樣品，其中血液樣品可以是血清或血漿樣品?？砂凑宅F(xiàn)有技術(shù)優(yōu)選地取得尿樣品。優(yōu)選地，中段尿樣品用于本發(fā)明上下文中所說的尿樣品。例如，還可利用WO01/74275中所述排尿設(shè)備收集尿樣品?？衫矛F(xiàn)有技術(shù)中7>知的方法取得血液樣品，例如，從靜脈、動脈或毛細管。通常，利用注射器例如從受試者的臂抽取靜脈血得到血液樣品。術(shù)語"血液樣品，，包括通過進一步純化和分離方法從血液中得到的樣品，比如血漿或血清?？衫矛F(xiàn)有技術(shù)中適于測量多肽標志物的任何公知方法測定樣品中多肽的存在與否。這樣的方法是技術(shù)人員公知的。原則上，可通過直接方法比如質(zhì)鐠、或間接方法例如通過配體測定多肽標志物的存在與否。如果有必要或有需要，可在測量多肽標志物存在與否之前通過任何適當?shù)姆椒A處理以及例如純化或分離來自受試者的樣品，例如尿或血液樣品。所述處理可包含例如純化、分離、稀釋或濃縮。所述方法可以是例如離心、過濾、超濾、透析、沉淀或色譜法，比如親和分離或利用離子交換色鐠的分離、電泳分離(即施加電場利用溶液中荷電顆粒的不同遷移行為進行分離)進行分離。其具體實例是凝膠電泳、二維聚丙烯酰胺凝膠電泳(2D-PAGE)、毛細管電泳、金屬親和色譜、固定化金屬親和色鐠(IMAC)、基于外源凝集素的親和色譜、液相色譜、高效液相色鐠(HPLC)、正相和反相HPLC、陽離子交換色譜以及選擇性表面結(jié)合。所有這些方法是技術(shù)人員熟知的，并且技術(shù)人員將能選擇用作所用樣品函數(shù)的方法以及選擇用于測定多肽標志物存在與否的方法。在本發(fā)明的一個實施方案中，在利用毛細管電泳進行分離之前，利用超速離心分離、純化樣品和/或利用超濾將樣品分成級分，所述級分含有特定分子大小的多肽標志物。優(yōu)選地，質(zhì)譜法用于測定多肽標志物的存在與否，其中可從這樣的方法出發(fā)逆向進行樣品的純化或分離。與目前所用的方法相比，質(zhì)譜分析具有可通過單一分析測定樣品中許多(>100)多肽之濃度的優(yōu)點?？墒褂萌魏晤愋偷馁|(zhì)鐠。利用質(zhì)鐠，可能在復雜的混合物中檢測lOfmol的多肽標志物(即O.lng的10kD蛋白質(zhì))，按照常規(guī)，測量精度為約±0.01%。在質(zhì)譜儀中，離子形成單元與適當?shù)姆治鲈O(shè)備耦聯(lián)。例如電噴霧離子化(ESI)接口主要用于測量液體樣品中的離子，而MALDI(基質(zhì)輔助激光解吸附/離子化)用于測量來自基質(zhì)中結(jié)晶化樣品的離子。為了分析形成的離子，可使用例如四極桿、離子阱或飛行時間(TOF)分析儀器。在電噴霧離子化(ESI)中，存在于溶液中的分子被霧化，此外，在高壓(例如l至8kV)的影響下，首先形成荷電的小滴，所述小滴變得更小從溶劑中蒸發(fā)。最后，所謂的庫侖爆炸導致形成游離離子，然后可分析和檢測所述游離離子。在利用TOF進行離子分析中，施加特定加速電壓，該電壓賦予離子等量的動能。其后，非常精確地測量各個離子通過飛行管移動特定漂移距離所需時間。由于具有等量的動能，離子的速度取決于它們的質(zhì)量，因此可檢測它們的質(zhì)量(后者)。TOF分析儀具有非常高的掃描速度并由此達到好的分辨率。用于檢測多肽標志物存在與否的優(yōu)選方法包括氣相離子色譜，比如激光解吸附/離子化質(zhì)鐠、MALDI-TOFMS、SELDI-TOFMS(表面增強激光解吸附/離子化)、LCMS(液相色語/質(zhì)譜)、2D-PAGE/MS以及毛細管電泳-質(zhì)譜(CE-MS)。上述所有方法是技術(shù)人員公知的。特別優(yōu)選的方法是CE-MS，其中毛細管電泳與質(zhì)語耦聯(lián)。該方法已經(jīng)相當詳細地被描述于例如德國專利申請DE10021737、Kaiser等(J.ChromatogrA，2003，Vol.l013:157-171，以及Electrophoresis,2004,25:2044-2055)、以及Wittke等(J.ChromatogrA,2003,1013:173-181)中。CE-MS技術(shù)允許在短時間內(nèi)和小體積中高靈敏度地同時測定樣品中數(shù)百種多肽標志物的存在。測量一種樣品后，制備所測量多肽標志物的模式(pattern),并且該模式可與患者或健康受試者的參比模式相比較。在多數(shù)情況下，使用有限數(shù)的多肽標志物用于UAS的診斷是足夠的。包含與ESI-TOFMS在線耦聯(lián)之CE的CE-MS法是進一步優(yōu)選的。對于CE-MS，使用揮發(fā)性溶劑是優(yōu)選的，并且最好在基本上無鹽條件下進行工作。這些溶劑的實例包括乙腈、異丙醇、甲醇等。所述溶劑可用水或弱酸(例如0.1%至1%的甲酸)稀釋以將分析物(優(yōu)選多肽)質(zhì)子化。利用毛細管電泳，可以通過分子的電荷和大小分離所述分子。施加電流后，中性粒子將以電滲流速度遷移，而陽離子加速朝向陽極，陰離子則被延遲。電泳中毛細管的優(yōu)點是保留有利的表面:體積比，其能在電流期間良好地消散產(chǎn)生的焦耳熱。從而允許施加高電壓(通常高達30kV)并因此允許高分離行為以及短的分析時間。在毛細管電泳中，通常4吏用內(nèi)徑典型為5075nm的珪玻璃毛細管。所用的長度為例如30100cm。此外，分離毛細管通常由涂敷塑料的硅玻璃制成。毛細管可以是未處理的(即它們的親水基團暴露于內(nèi)表面上)或者在內(nèi)表面上有涂層。疏水性涂層可用于提高分辨率。除了電壓以外，還可施加壓力，所加壓力通常在0至lpsi的范圍內(nèi)。還可僅僅在分離或改變期間同時施加壓力。在一個優(yōu)選的用于測量多肽標志物的方法中，利用毛細管電泳分離樣品的標志物，然后直接離子化并在線轉(zhuǎn)移到耦聯(lián)的質(zhì)鐠儀中用于檢測。在根據(jù)本發(fā)明的方法中，使用幾種多肽標志物用于診斷VD是有利的。具體而言，至少可使用三種多肽標志物，例如標志物l、2和3;1、2和4;等等。使用至少4、5或6種標志物是更優(yōu)選的。使用至少ll種標志物，例如標志物l至ll，甚至是更優(yōu)選的。使用表l至4中所列舉的所有526種標志物是最優(yōu)選的。為了在使用幾種標志物時測定存在嚴重VD的概率，可使用技術(shù)人員公知的統(tǒng)計學方法。例如，通過利用計算機程序比如S-Plus,或者Weissinger等(KidneyInt.,2004,65:2426-2434)所述的支持向量機，可使用上述同一文獻中所述的RandomForests法。實施例1.樣品制備對于檢測多肽標志物以i貪斷VD，以及患有嚴重VD的患者收集尿。使用尿。從健康供體(對照組)對于隨后的CE-MS檢測，必須通過超濾分離掉以高濃度同樣包含于患者尿中的蛋白質(zhì)(比如白蛋白和免疫球蛋白)。因此，收集700pJ尿并與700nm的過濾緩沖液(2M尿素、10mM氨、0.02。/。SDS)相混合。超濾該1.4ml體積的樣品(20kDa，SartoriusG6ttingen，德國)。在離心機中3000rpm下進行所述超濾直到得到l.lml超濾液。然后將所得的l.lml超濾液施加到PD10柱(AmershamBioscience,Uppsala，瑞典)并用2.5ml0.01。/。NH40H洗脫，并凍干。為了CE-MS檢測，然后用20nl7jC(HPLC級，Merck)重懸所述多肽。2.CE-MS檢測利用BeckmanCoulter的毛細管電泳系統(tǒng)(P/ACEMDQSystem;BeckmanCoulterInc.,Fullerton,CA,USA)和Bruker的ESI國TOF質(zhì)鐠儀(micro-TOFMS,BrukerDaltonik,Bremen,德國)進行CE畫MS檢測。CE毛細管由BeckmanCoulter提供并且ID/OD為50/360|Lim、長度為90cm。CE分離的流動相由含20%乙腈和0.25%甲酸的水組成。對于MS上的"鞘流"，使用含0.5%甲酸的30%異丙醇，此處的流速為2pl/分鐘。CE和MS的耦聯(lián)通過CE-ESI-MSSprayer套盒(AgilentTechnologies,Waldbronn，德國)實現(xiàn)。對于注射樣品，施用1至最大6psi的壓力，并且持續(xù)注射時間為99秒。利用這些參數(shù)，將約150nl樣品注射到毛細管中，其相當于毛細管體積的約10%。在毛細管中用堆疊技術(shù)(stackingtechnique)濃縮樣品。因此，注射才羊品之前，注射1MNH3溶液7秒(lpsi下)，并在注射樣品后，注射2M甲酸溶液5秒。當施加分離電壓(30kV)時，分析物在這些溶液之間被自動濃縮。利用壓力法進4亍隨后的CE分離0psi40分鐘，然后0.1psi2分鐘，0.2psi2分鐘，0.3psi2分鐘,0.4psi2分鐘，最后0.5psi32分鐘。因此分離操作的總持續(xù)時間為80分鐘。為了在MS—側(cè)得到盡可能好的信號強度，霧化氣體被設(shè)置至最低可能的值。施加到噴針用于產(chǎn)生電噴霧的電壓是3700~4100V。根據(jù)廠商的說明書對質(zhì)i普儀上的其余設(shè)置進行最優(yōu)化以進行肽檢測。在m/z400至m/z3000質(zhì)量范圍內(nèi)記錄鐠圖并每3秒鐘進行累計。3.用于CE檢測的標準對于檢驗和標準化CE檢測，使用特征在于所述CE遷移時間的下述蛋白質(zhì)或多肽蛋白質(zhì)/多肽遷移時間抑肽酶(SIGMA,Taufkirchen,DE,Cat.#AI153)9.2分鐘核糖核酸酶(SIGMA,Taufkirchen,DE,Cat.#R487S)10,9分鐘溶菌酶(SIGMA,Taufkirchen,DE,Cat.#L7651)8.9分鐘"REV",序列REVQSKIGYGRQIIS15.6分鐘"ELM",序列ELMTGELPYSHINNRDQIIFMVGR23.4分,中"KINCON",序列TGSLPYSHIGSRDQIIFMVGR20.0分4中"GIVLY"序列GIVLYELMTGELPYSHIN36.8分鐘以在水中10pmol/pl的濃度使用每種蛋白質(zhì)/多肽。"REV，，、"ELM"、"KINCON"和"GIVLY"是合成肽。肽的分子質(zhì)量和在MS中可見的單個電荷態(tài)之m/z比被描述于下述表中單<table>tableseeoriginaldocumentpage27</column></row><table>原則上，在利用毛細管電泳的分離中可能發(fā)生遷移時間的輕微變化是技術(shù)人員公知的。然而，在所述條件下，遷移順序?qū)⒉粫淖儭τ谥浪鲑|(zhì)量和CE時間的^t術(shù)人員，可以毫不費力地將他們自己的測定分配給根據(jù)本發(fā)明的多肽標志物。例如，他可按照下述進行首先，他選擇一種在他的測量中發(fā)現(xiàn)的多肽(肽1)并試圖在所述CE時間的時隙內(nèi)(例如士5分鐘)發(fā)現(xiàn)一種或多種相同的質(zhì)量。如果在此間隔內(nèi)僅僅發(fā)現(xiàn)一種相同的質(zhì)量，則分配完成。如果發(fā)現(xiàn)數(shù)種匹配的質(zhì)量，有關(guān)該分配的決定仍然需要進行。因此，從檢測中選擇另外一種肽(肽2),并努力確認適當?shù)碾臉酥疚?，再次考慮相應(yīng)的時隙。此外，如果可發(fā)現(xiàn)數(shù)種標志物具有對應(yīng)的質(zhì)量，最可能的分配是其中肽l的遷移和肽2的遷移之間具有基本上線性的關(guān)系。根據(jù)分配問題的復雜性，建iU支術(shù)人員自身任選地使用來自其樣品的其它蛋白質(zhì)用于分配，例如十種蛋白質(zhì)。通常，遷移時間延長或縮短特定的絕對值，或者出現(xiàn)整個過程的壓縮或延伸。然而，在這樣的條件下共遷移肽也將共遷移。此外，技術(shù)人員可利用Zuerbig等在Electrophoresis27(2006)，pp.2111-2125中所述的遷移模式。如果他利用筒^圖表(例如利用MSExcel)以m/z相對于遷移時間的形式對其測量進行作圖，所述的線模式也變得明顯?，F(xiàn)在，通過線計數(shù)使得可以簡單地分配單個多肽。也可以使用其它的分配方法。基本地，技術(shù)人員還可使用上述肽作為內(nèi)標用于分配他的CE測量。權(quán)利要求1.一種診斷血管病(VD)的方法，其包括測定樣品中是否存在至少一種多肽標志物的步驟，其中所述多肽標志物選自標志物1～526，其特征在于具有下述的分子質(zhì)量和遷移時間<tablesid="tabl0001"num="0001"><table><tgroupcols="15"><colspeccolname="c001"colwidth="5%"/><colspeccolname="c002"colwidth="8%"/><colspeccolname="c003"colwidth="8%"/><colspeccolname="c004"colwidth="5%"/><colspeccolname="c005"colwidth="7%"/><colspeccolname="c006"colwidth="8%"/><colspeccolname="c007"colwidth="5%"/><colspeccolname="c008"colwidth="7%"/><colspeccolname="c009"colwidth="8%"/><colspeccolname="c010"colwidth="4%"/><colspeccolname="c011"colwidth="7%"/><colspeccolname="c012"colwidth="8%"/><colspeccolname="c013"colwidth="4%"/><colspeccolname="c014"colwidth="8%"/><colspeccolname="c015"colwidth="8%"/><thead></column></row><row><column><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry></column></row></thead><tbody></column></row><row><column><entrymorerows="1">1</entry><entrymorerows="1">1166.61</entry><entrymorerows="1">23.88</entry><entrymorerows="1">51</entry><entrymorerows="1">11058.18</entry><entrymorerows="1">21.88</entry><entrymorerows="1">101</entry><entrymorerows="1">3984.81</entry><entrymorerows="1">21.29</entry><entrymorerows="1">151</entry><entrymorerows="1">2082.01</entry><entrymorerows="1">33.67</entry><entrymorerows="1">201</entry><entrymorerows="1">1013.41</entry><entrymorerows="1">25.33</entry></column></row></column></row><row><column><entrymorerows="1">2</entry><entrymorerows="1">2431.50</entry><entrymorerows="1">24.10</entry><entrymorerows="1">52</entry><entrymorerows="1">1352.61</entry><entrymorerows="1">29.86</entry><entrymorerows="1">102</entry><entrymorerows="1">4830.78</entry><entrymorerows="1">26.61</entry><entrymorerows="1">152</entry><entrymorerows="1">1768.90</entry><entrymorerows="1">20.77</entry><entrymorerows="1">202</entry><entrymorerows="1">1016.49</entry><entrymorerows="1">25.88</entry></column></row></column></row><row><column><entrymorerows="1">3</entry><entrymorerows="1">1922.93</entry><entrymorerows="1">31.99</entry><entrymorerows="1">53</entry><entrymorerows="1">2802.85</entry><entrymorerows="1">36.35</entry><entrymorerows="1">103</entry><entrymorerows="1">3031.39</entry><entrymorerows="1">35.93</entry><entrymorerows="1">153</entry><entrymorerows="1">3442.09</entry><entrymorerows="1">33.32</entry><entrymorerows="1">203</entry><entrymorerows="1">1493.74</entry><entrymorerows="1">22.06</entry></column></row></column></row><row><column><entrymorerows="1">4</entry><entrymorerows="1">2509.16</entry><entrymorerows="1">25.76</entry><entrymorerows="1">54</entry><entrymorerows="1">4890.88</entry><entrymorerows="1">26.46</entry><entrymorerows="1">104</entry><entrymorerows="1">3788.76</entry><entrymorerows="1">25.21</entry><entrymorerows="1">154</entry><entrymorerows="1">876.42</entry><entrymorerows="1">35.07</entry><entrymorerows="1">204</entry><entrymorerows="1">2104.04</entry><entrymorerows="1">32.97</entry></column></row></column></row><row><column><entrymorerows="1">5</entry><entrymorerows="1">3194.22</entry><entrymorerows="1">30.34</entry><entrymorerows="1">55</entry><entrymorerows="1">5212.06</entry><entrymorerows="1">26.98</entry><entrymorerows="1">105</entry><entrymorerows="1">2567.2</entry><entrymorerows="1">34.76</entry><entrymorerows="1">155</entry><entrymorerows="1">2352.14</entry><entr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758.37</entry><entrymorerows="1">28.94</entry><entrymorerows="1">94</entry><entrymorerows="1">3303.00</entry><entrymorerows="1">23.07</entry><entrymorerows="1">144</entry><entrymorerows="1">1898.93</entry><entrymorerows="1">40.30</entry><entrymorerows="1">194</entry><entrymorerows="1">3489.70</entry><entrymorerows="1">31.45</entry><entrymorerows="1">244</entry><entrymorerows="1">1608.76</entry><entrymorerows="1">22.36</entry></column></row></column></row><row><column><entrymorerows="1">45</entry><entrymorerows="1">3546.94</entry><entrymorerows="1">26.22</entry><entrymorerows="1">95</entry><entrymorerows="1">4081.56</entry><entrymorerows="1">24.51</entry><entrymorerows="1">145</entry><entrymorerows="1">2237.06</entry><entrymorerows="1">27.12</entry><entrymorerows="1">195</entry><entrymorerows="1">1268.62</entry><entrymorerows="1">27.29</entry><entrymorerows="1">245</entry><entrymorerows="1">2501.20</entry><entrymorerows="1">34.30</entry></column></row></column></row><row><column><entrymorerows="1">46</entry><entrymorerows="1">3609.63</entry><entrymorerows="1">20.22</entry><entrymorerows="1">96</entry><entrymorerows="1">4670.27</entry><entrymorerows="1">25.84</entry><entrymorerows="1">146</entry><entrymorerows="1">3178.33</entry><entrymorerows="1">30.26</entry><entrymorerows="1">196</entry><entrymorerows="1">1659.82</entry><entrymorerows="1">29.35</entry><entrymorerows="1">246</entry><entrymorerows="1">3021.52</entry><entrymorerows="1">23.52</entry></column></row></column></row><row><column><entrymorerows="1">47</entry><entrymorerows="1">3697.49</entry><entrymorerows="1">23.71</entry><entrymorerows="1">97</entry><entrymorerows="1">4671.99</entry><entrymorerows="1">23.33</entry><entrymorerows="1">147</entry><entrymorerows="1">1844.56</entry><entrymorerows="1">34.28</entry><entrymorerows="1">197</entry><entrymorerows="1">2405.59</entry><entrymorerows="1">22.16</entry><entrymorerows="1">247</entry><entrymorerows="1">4153.75</entry><entrymorerows="1">33.41</entry></column></row></column></row><row><column><entrymorerows="1">48</entry><entrymorerows="1">4278.73</entry><entrymorerows="1">23.34</entry><entrymorerows="1">98</entry><entrymorerows="1">8933.94</entry><entrymorerows="1">22.57</entry><entrymorerows="1">148</entry><entrymorerows="1">1378.57</entry><entrymorerows="1">37.16</entry><entrymorerows="1">198</entry><entrymorerows="1">2483.21</entry><entrymorerows="1">27.54</entry><entrymorerows="1">248</entry><entrymorerows="1">5000.17</entry><entrymorerows="1">24.43</entry></column></row></column></row><row><column><entrymorerows="1">49</entry><entrymorerows="1">4421.04</entry><entrymorerows="1">20.73</entry><entrymorerows="1">99</entry><entrymorerows="1">1523.90</entry><entrymorerows="1">29.72</entry><entrymorerows="1">149</entry><entrymorerows="1">1764.86</entry><entrymorerows="1">29.88</entry><entrymorerows="1">199</entry><entrymorerows="1">2599.21</entry><entrymorerows="1">28.20</entry><entrymorerows="1">249</entry><entrymorerows="1">8917.48</entry><entrymorerows="1">22.53</entry></column></row></column></row><row><column><entrymorerows="1">50</entry><entrymorerows="1">4805.67</entry><entrymorerows="1">26.49</entry><entrymorerows="1">100</entry><entrymorerows="1">3956.82</entry><entrymorerows="1">25.20</entry><entrymorerows="1">150</entry><entrymorerows="1">1791.88</entry><entrymorerows="1">30.77</entry><entrymorerows="1">200</entry><entrymorerows="1">4170.01</entry><entrymorerows="1">33.51</entry><entrymorerows="1">250</entry><entrymorerows="1">1750.86</entry><entrymorerows="1">23.80</entry></column></row></tbody></tgroup></column></row><table></tables><tablesid="tabl0002"num="0002"><table><tgroupcols="12"><colspeccolname="c001"colwidth="6%"/><colspeccolname="c002"colwidth="10%"/><colspeccolname="c003"colwidth="10%"/><colspeccolname="c004"colwidth="6%"/><colspeccolname="c005"colwidth="9%"/><colspeccolname="c006"colwidth="10%"/><colspeccolname="c007"colwidth="6%"/><colspeccolname="c008"colwidth="9%"/><colspeccolname="c009"colwidth="9%"/><colspeccolname="c010"colwidth="6%"/><colspeccolname="c011"colwidth="9%"/><colspeccolname="c012"colwidth="10%"/><thead></column></row><row><column><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry></column></row></thead><tbody></column></row><row><column><entrymorerows="1">251</entry><entrymorerows="1">2235.13</entry><entrymorerows="1">34.10</entry><entrymorerows="1">301</entry><entrymorerows="1">1649.79</entry><entrymorerows="1">19.59</entry><entrymorerows="1">351</entry><entrymorerows="1">2073.17</entry><entrymorerows="1">27.43</entry><entrymorerows="1">401</entry><entrymorerows="1">1749.88</entry><entrymorerows="1">30.54</entry></column></row></column></row><row><column><entrymorerows="1">252</entry><entrymorerows="1">2644.25</entry><entrymorerows="1">21.13</entry><entrymorerows="1">302</entry><entrymorerows="1">4025.68</entry><entrymorerows="1">20.73</entry><entrymorerows="1">352</entry><entrymorerows="1">7958.65</entry><entrymorerows="1">34.32</entry><entrymorerows="1">402</entry><entrymorerows="1">1956.97</entry><entrymorerows="1">21.44</entry></column></row></column></row><row><column><entrymorerows="1">253</entry><entrymorerows="1">3943.96</entry><entrymorerows="1">33.53</entry><entrymorerows="1">303</entry><entrymorerows="1">980.33</entry><entrymorerows="1">35.59</entry><entrymorerows="1">353</entry><entrymorerows="1">1837.88</entry><entrymorerows="1">30.54</entry><entrymorerows="1">403</entry><entrymorerows="1">2189.12</entry><entrymorerows="1">26.54</entry></column></row></column></row><row><column><entrymorerows="1">254</entry><entrymorerows="1">11967.96</entry><entrymorerows="1">20.50</entry><entrymorerows="1">304</entry><entrymorerows="1">1096.41</entry><entrymorerows="1">35.95</entry><entrymorerows="1">354</entry><entrymorerows="1">2939.03</entry><entrymorerows="1">33.75</entry><entrymorerows="1">404</entry><entrymorerows="1">2257.63</entry><entrymorerows="1">36.10</entry></column></row></column></row><row><column><entrymorerows="1">255</entry><entrymorerows="1">2553.23</entry><entrymorerows="1">34.14</entry><entrymorerows="1">305</entry><entrymorerows="1">1698.65</entry><entrymorerows="1">37.60</entry><entrymorerows="1">355</entry><entrymorerows="1">2977.31</entry><entrymorerows="1">19.59</entry><entrymorerows="1">405</entry><entrymorerows="1">2917.54</entry><entrymorerows="1">28.99</entry></column></row></column></row><row><column><entrymorerows="1">256</entry><entrymorerows="1">1209.58</entry><entrymorerows="1">26.31</entry><entrymorerows="1">306</entry><entrymorerows="1">2361.21</entry><entrymorerows="1">20.77</entry><entrymorerows="1">356</entry><entrymorerows="1">3596.46</entry><entrymorerows="1">21.54</entry><entrymorerows="1">406</entry><entrymorerows="1">3633.69</entry><entrymorerows="1">26.99</entry></column></row></column></row><row><column><entrymorerows="1">257</entry><entrymorerows="1">1899.94</entry><entrymorerows="1">21.41</entry><entrymorerows="1">307</entry><entrymorerows="1">3148.50</entry><entrymorerows="1">24.22</entry><entrymorerows="1">357</entry><entrymorerows="1">3851.68</entry><entrymorerows="1">24.97</entry><entrymorerows="1">407</entry><entrymorerows="1">6055.77</entry><entrymorerows="1">21.03</entry></column></row></column></row><row><column><entrymorerows="1">258</entry><entrymorerows="1">1680.00</entry><entrymorerows="1">23.77</entry><entrymorerows="1">308</entry><entrymorerows="1">3157.23</entry><entrymorerows="1">34.74</entry><entrymorerows="1">358</entry><entrymorerows="1">1135.52</entry><entrymorerows="1">27.83</entry><entrymorerows="1">408</entry><entrymorerows="1">6186.02</entry><entrymorerows="1">24.99</entry></column></row></column></row><row><column><entrymorerows="1">259</entry><entrymorerows="1">2195.06</entry><entrymorerows="1">20.15</entry><entrymorerows="1">309</entry><entrymorerows="1">1304.59</entry><entrymorerows="1">27.95</entry><entrymorerows="1">359</entry><entrymorerows="1">3378.05</entry><entrymorerows="1">38.81</entry><entrymorerows="1">409</entry><entrymorerows="1">1858.92</entry><entrymorerows="1">24.17</entry></column></row></column></row><row><column><entrymorerows="1">260</entry><entrymorerows="1">3064.41</entry><entrymorerows="1">20.55</entry><entrymorerows="1">310</entry><entrymorerows="1">3575.78</entry><entrymorerows="1">32.27</entry><entrymorerows="1">360</entry><entrymorerows="1">3590.72</entry><entrymorerows="1">28.99</entry><entrymorerows="1">410</entry><entrymorerows="1">2274.11</entry><entrymorerows="1">33.47</entry></column></row></column></row><row><column><entrymorerows="1">261</entry><entrymorerows="1">3554.07</entry><entrymorerows="1">31.11</entry><entrymorerows="1">311</entry><entrymorerows="1">1510.75</entry><entrymorerows="1">20.12</entry><entrymorerows="1">361</entry><entrymorerows="1">3959.80</entry><entrymorerows="1">19.95</entry><entrymorerows="1">411</entry><entrymorerows="1">4522.51</entry><entrymorerows="1">26.20</entry></column></row></column></row><row><column><entrymorerows="1">262</entry><entrymorerows="1">3686.03</entry><entrymorerows="1">22.16</entry><entrymorerows="1">312</entry><entrymorerows="1">2485.20</entry><entrymorerows="1">34.25</entry><entrymorerows="1">352</entry><entrymorerows="1">1258.41</entry><entrymorerows="1">36.10</entry><entrymorerows="1">412</entry><entrymorerows="1">6237.35</entry><entrymorerows="1">31.39</entry></column></row></column></row><row><column><entrymorerows="1">263</entry><entrymorerows="1">3802.12</entry><entrymorerows="1">33.10</entry><entrymorerows="1">313</entry><entrymorerows="1">3076.33</entry><entrymorerows="1">19.64</entry><entrymorerows="1">363</entry><entrymorerows="1">1513.50</entry><entrymorerows="1">36.82<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="1">33.26</entry><entrymorerows="1">375</entry><entrymorerows="1">2777.71</entry><entrymorerows="1">21.55</entry><entrymorerows="1">425</entry><entrymorerows="1">1635.8</entry><entrymorerows="1">40.33</entry></column></row></column></row><row><column><entrymorerows="1">276</entry><entrymorerows="1">1846.93</entry><entrymorerows="1">32.04</entry><entrymorerows="1">326</entry><entrymorerows="1">2411.78</entry><entrymorerows="1">26.97</entry><entrymorerows="1">376</entry><entrymorerows="1">3521.02</entry><entrymorerows="1">30.73</entry><entrymorerows="1">426</entry><entrymorerows="1">2525.2</entry><entrymorerows="1">27.72</entry></column></row></column></row><row><column><entrymorerows="1">277</entry><entrymorerows="1">3177.14</entry><entrymorerows="1">22.48</entry><entrymorerows="1">327</entry><entrymorerows="1">3738.59</entry><entrymorerows="1">24.76</entry><entrymorerows="1">377</entry><entrymorerows="1">3750.72</entry><entrymorerows="1">32.45</entry><entrymorerows="1">427</entry><entrymorerows="1">1526.8</entry><entrymorerows="1">23.63</entry></column></row></column></row><row><column><entrymorerows="1">278</entry><entrymorerows="1">4113.80</entry><entrymorerows="1">24.58</entry><entrymorerows="1">328</entry><entrymorerows="1">3935.57</entry><entrymorerows="1">34.15</entry><entrymorerows="1">378</entry><entrymorerows="1">4229.09</entry><entrymorerows="1">29.08</entry><entrymorerows="1">428</entry><entrymorerows="1">1664.8</entry><entrymorerows="1">29.87</entry></column></row></column></row><row><column><entrymorerows="1">279</entry><entrymorerows="1">2744.07</entry><entrymorerows="1">35.03</entry><entrymorerows="1">329</entry><entrymorerows="1">4863.21</entry><entrymorerows="1">26.66</entry><entrymorerows="1">379</entry><entrymorerows="1">4846.50</entry><entrymorerows="1">26.40</entry><entrymorerows="1">429</entry><entrymorerows="1">2583.3</entry><entrymorerows="1">28.31</entry></column></row></column></row><row><column><entrymorerows="1">280</entry><entrymorerows="1">2767.26</entry><entrymorerows="1">21.52</entry><entrymorerows="1">330</entry><entrymorerows="1">860.39</entry><entrymorerows="1">26.25</entry><entrymorerows="1">380</entry><entrymorerows="1">1046.55</entry><entrymorerows="1">25.35</entry><entrymorerows="1">430</entry><entrymorerows="1">2663.3</entry><entrymorerows="1">23.44</entry></column></row></column></row><row><column><entrymorerows="1">281</entry><entrymorerows="1">1310.64</entry><entrymorerows="1">27.11</entry><entrymorerows="1">331</entry><entrymorerows="1">1567.78</entry><entrymorerows="1">20.23</entry><entrymorerows="1">381</entry><entrymorerows="1">1608.80</entry><entrymorerows="1">30.94</entry><entrymorerows="1">431</entry><entrymorerows="1">1878.9</entry><entrymorerows="1">42.18</entry></column></row></column></row><row><column><entrymorerows="1">282</entry><entrymorerows="1">1613.88</entry><entrymorerows="1">23.95</entry><entrymorerows="1">332</entry><entrymorerows="1">2308.11</entry><entrymorerows="1">27.32</entry><entrymorerows="1">382</entry><entrymorerows="1">1878.78</entry><entrymorerows="1">31.58</entry><entrymorerows="1">432</entry><entrymorerows="1">1462.7</entry><entrymorerows="1">39.31</entry></column></row></column></row><row><column><entrymorerows="1">283</entry><entrymorerows="1">1703.90</entry><entrymorerows="1">33.64</entry><entrymorerows="1">333</entry><entrymorerows="1">2923.77</entry><entrymorerows="1">36.44</entry><entrymorerows="1">383</entry><entrymorerows="1">2589.16</entry><entrymorerows="1">22.45</entry><entrymorerows="1">433</entry><entrymorerows="1">1834.9</entry><entrymorerows="1">24</entry></column></row></column></row><row><column><entrymorerows="1">284</entry><entrymorerows="1">2761.40</entry><entrymorerows="1">21.46</entry><entrymorerows="1">334</entry><entrymorerows="1">3295.55</entry><entrymorerows="1">25.40</entry><entrymorerows="1">384</entry><entrymorerows="1">4369.06</entry><entrymorerows="1">20.25</entry><entrymorerows="1">434</entry><entrymorerows="1">1893.1</entry><entrymorerows="1">24.64</entry></column></row></column></row><row><column><entrymorerows="1">285</entry><entrymorerows="1">3242.42</entry><entrymorerows="1">22.78</entry><entrymorerows="1">335</entry><entrymorerows="1">3870.85</entry><entrymorerows="1">33.39</entry><entrymorerows="1">385</entry><entrymorerows="1">12717.08</entry><entrymorerows="1">26.92</entry><entrymorerows="1">435</entry><entrymorerows="1">1934</entry><entrymorerows="1">21.63</entry></column></row></column></row><row><column><entrymorerows="1">286</entry><entrymorerows="1">3338.17</entry><entrymorerows="1">23.36</entry><entrymorerows="1">336</entry><entrymorerows="1">1099.56</entry><entrymorerows="1">21.63</entry><entrymorerows="1">386</entry><entrymorerows="1">1210.43</entry><entrymorerows="1">36.48</entry><entrymorerows="1">436</entry><entrymorerows="1">1367.7</entry><entrymorerows="1">38.87</entry></column></row></column></row><row><column><entrymorerows="1">287</entry><entrymorerows="1">3371.74</entry><entrymorerows="1">22.96</entry><entrymorerows="1">337</entry><entrymorerows="1">1359.70</entry><entrymorerows="1">22.92</entry><entrymorerows="1">387</entry><entrymorerows="1">3092.54</entry><entrymorerows="1">36.22</entry><entrymorerows="1">437</entry><entrymorerows="1">1009.5</entry><entrymorerows="1">27.33</entry></column></row></column></row><row><column><entrymorerows="1">288</entry><entrymorerows="1">3593.53</entry><entrymorerows="1">20.25</entry><entrymorerows="1">338</entry><entrymorerows="1">2059.02</entry><entrymorerows="1">23.12</entry><entrymorerows="1">388</entry><entrymorerows="1">3246.61</entry><entrymorerows="1">25.65</entry><entrymorerows="1">438</entry><entrymorerows="1">3405.1</entry><entrymorerows="1">25.92</entry></column></row></column></row><row><column><entrymorerows="1">289</entry><entrymorerows="1">3677.52</entry><entrymorerows="1">24.49</entry><entrymorerows="1">339</entry><entrymorerows="1">2077.03</entry><entrymorerows="1">21.78</entry><entrymorerows="1">389</entry><entrymorerows="1">4012.41</entry><entrymorerows="1">20.81</entry><entrymorerows="1">439</entry><entrymorerows="1">2314.1</entry><entrymorerows="1">33.67</entry></column></row></column></row><row><column><entrymorerows="1">290</entry><entrymorerows="1">1624.80</entry><entrymorerows="1">30.81</entry><entrymorerows="1">340</entry><entrymorerows="1">3349.34</entry><entrymorerows="1">35.81</entry><entrymorerows="1">390</entry><entrymorerows="1">11016.34</entry><entrymorerows="1">21.31</entry><entrymorerows="1">440</entry><entrymorerows="1">3996.8</entry><entrymorerows="1">20.93</entry></column></row></column></row><row><column><entrymorerows="1">291</entry><entrymorerows="1">2210.92</entry><entrymorerows="1">37.55</entry><entrymorerows="1">341</entry><entrymorerows="1">8853.85</entry><entrymorerows="1">21.08</entry><entrymorerows="1">391</entry><entrymorerows="1">1284.61</entry><entrymorerows="1">29.17</entry><entrymorerows="1">441</entry><entrymorerows="1">2823.6</entry><entrymorerows="1">29.07</entry></column></row></column></row><row><column><entrymorerows="1">292</entry><entrymorerows="1">3290.37</entry><entrymorerows="1">24.12</entry><entrymorerows="1">342</entry><entrymorerows="1">1734.80</entry><entrymorerows="1">20.24</entry><entrymorerows="1">392</entry><entrymorerows="1">1460.83</entry><entrymorerows="1">22.53</entry><entrymorerows="1">442</entry><entrymorerows="1">1179.6</entry><entrymorerows="1">27.15</entry></column></row></column></row><row><column><entrymorerows="1">293</entry><entrymorerows="1">4413.76</entry><entrymorerows="1">29.03</entry><entrymorerows="1">343</entry><entrymorerows="1">1847.95</entry><entrymorerows="1">43.93</entry><entrymorerows="1">393</entry><entrymorerows="1">1807.88</entry><entrymorerows="1">23.98</entry><entrymorerows="1">443</entry><entrymorerows="1">1435.7</entry><entrymorerows="1">28.86</entry></column></row></column></row><row><column><entrymorerows="1">294</entry><entrymorerows="1">1482.73</entry><entrymorerows="1">22.47</entry><entrymorerows="1">344</entry><entrymorerows="1">2045.95</entry><entrymorerows="1">34.04</entry><entrymorerows="1">394</entry><entrymorerows="1">2596.33</entry><entrymorerows="1">34.86</entry><entrymorerows="1">444</entry><entrymorerows="1">2430.7</entry><entrymorerows="1">35.39</entry></column></row></column></row><row><column><entrymorerows="1">295</entry><entrymorerows="1">1813.78</entry><entrymorerows="1">31.87</entry><entrymorerows="1">345</entry><entrymorerows="1">2289.47</entry><entrymorerows="1">33.56</entry><entrymorerows="1">395</entry><entrymorerows="1">2686.97</entry><entrymorerows="1">29.06</entry><entrymorerows="1">445</entry><entrymorerows="1">1134.6</entry><entrymorerows="1">23.68</entry></column></row></column></row><row><column><entrymorerows="1">296</entry><entrymorerows="1">1934.87</entry><entrymorerows="1">20.04</entry><entrymorerows="1">346</entry><entrymorerows="1">2421.15</entry><entrymorerows="1">34.74</entry><entrymorerows="1">396</entry><entrymorerows="1">3871.59</entry><entrymorerows="1">27.51</entry><entrymorerows="1">446</entry><entrymorerows="1">2014</entry><entrymorerows="1">25.18</entry></column></row></column></row><row><column><entrymorerows="1">297</entry><entrymorerows="1">2249.89</entry><entrymorerows="1">34.14</entry><entrymorerows="1">347</entry><entrymorerows="1">2480.67</entry><entrymorerows="1">23.00</entry><entrymorerows="1">397</entry><entrymorerows="1">4069.63</entry><entrymorerows="1">25.30</entry><entrymorerows="1">447</entry><entrymorerows="1">2577.3</entry><entrymorerows="1">24.55</entry></column></row></column></row><row><column><entrymorerows="1">298</entry><entrymorerows="1">3280.59</entry><entrymorerows="1">25.76</entry><entrymorerows="1">348</entry><entrymorerows="1">2576.25</entry><entrymorerows="1">34.17</entry><entrymorerows="1">398</entry><entrymorerows="1">4288.98</entry><entrymorerows="1">25.94</entry><entrymorerows="1">448</entry><entrymorerows="1">1194.6</entry><entrymorerows="1">26.73</entry></column></row></column></row><row><column><entrymorerows="1">299</entry><entrymorerows="1">1096.56</entry><entrymorerows="1">21.46</entry><entrymorerows="1">349</entry><entrymorerows="1">3353.93</entry><entrymorerows="1">23.53</entry><entrymorerows="1">399</entry><entrymorerows="1">4426.21</entry><entrymorerows="1">20.09</entry><entrymorerows="1">449</entry><entrymorerows="1">1588.8</entry><entrymorerows="1">30.2</entry></column></row></column></row><row><column><entrymorerows="1">300</entry><entrymorerows="1">1125.58</entry><entrymorerows="1">21.76</entry><entrymorerows="1">350</entry><entrymorerows="1">1063.52</entry><entrymorerows="1">26.24</entry><entrymorerows="1">400</entry><entrymorerows="1">1071.55</entry><entrymorerows="1">21.41</entry><entrymorerows="1">450</entry><entrymorerows="1">2056</entry><entrymorerows="1">25.44</entry></column></row></tbody></tgroup></column></row><table></tables><tablesid="tabl0003"num="0003"><table><tgroupcols="6"><colspeccolname="c001"colwidth="12%"/><colspeccolname="c002"colwidth="19%"/><colspeccolname="c003"colwidth="19%"/><colspeccolname="c004"colwidth="11%"/><colspeccolname="c005"colwidth="19%"/><colspeccolname="c006"colwidth="20%"/><thead></column></row><row><column><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry><entrymorerows="1">No.</entry><entrymorerows="1">質(zhì)量</entry><entrymorerows="1">CE時間</entry></column></row></thead><tbody></column></row><row><column><entrymorerows="1">451</entry><entrymorerows="1">2442.16</entry><entrymorerows="1">34.11</entry><entrymorerows="1">501</entry><entrymorerows="1">1173.58</entry><entrymorerows="1">37.51</entry></column></row></column></row><row><column><entrymorerows="1">452</entry><entrymorerows="1">1422.66</entry><entrymorerows="1">21.72</entry><entrymorerows="1">502</entry><entrymorerows="1">1100.55</entry><entrymorerows="1">36.99</entry></column></row></column></row><row><column><entrymorerows="1">453</entry><entrymorerows="1">1623.8</entry><entrymorerows="1">24.15</entry><entrymorerows="1">503</entry><entrymorerows="1">1128.44</entry><entrymorerows="1">33.71</entry></column></row></column></row><row><column><entrymorerows="1">454</entry><entrymorerows="1">1624.61</entry><entrymorerows="1">37.73</entry><entrymorerows="1">504</entry><entrymorerows="1">3149.6</entry><entrymorerows="1">31.22</entry></column></row></column></row><row><column><entrymorerows="1">455</entry><entrymorerows="1">3298.48</entry><entrymorerows="1">36.06</entry><entrymorerows="1">505</entry><entrymorerows="1">1068.56</entry><entrymorerows="1">21.69</entry></column></row></column></row><row><column><entrymorerows="1">456</entry><entrymorerows="1">1016.31</entry><entrymorerows="1">35.67</entry><entrymorerows="1">506</entry><entrymorerows="1">1349.48</entry><entrymorerows="1">36.47</entry></column></row></column></row><row><column><entrymorerows="1">457</entry><entrymorerows="1">1580.94</entry><entrymorerows="1">24.31</entry><entrymorerows="1">507</entry><entrymorerows="1">1889.81</entry><entrymorerows="1">40.57</entry></column></row></column></row><row><column><entrymorerows="1">458</entry><entrymorerows="1">1157.58</entry><entrymorerows="1">37.41</entry><entrymorerows="1">508</entry><entrymorerows="1">2305.7</entry><entrymorerows="1">34.8</entry></column></row></column></row><row><column><entrymorerows="1">459</entry><entrymorerows="1">1250.61</entry><entrymorerows="1">27.94</entry><entrymorerows="1">509</entry><entrymorerows="1">840.44</entry><entrymorerows="1">23.94</entry></column></row></column></row><row><column><entrymorerows="1">460</entry><entrymorerows="1">1378.67</entry><entrymorerows="1">28.85</entry><entrymorerows="1">510</entry><entrymorerows="1">911.3</entry><entrymorerows="1">34.39</entry></column></row></column></row><row><column><entrymorerows="1">461</entry><entrymorerows="1">1392.68</entry><entrymorerows="1">21.75</entry><entrymorerows="1">511</entry><entrymorerows="1">1299.64</entry><entrymorerows="1">22.42</entry></column></row></column></row><row><column><entrymorerows="1">462</entry><entrymorerows="1">1409.64</entry><entrymorerows="1">22.06</entry><entrymorerows="1">512</entry><entrymorerows="1">911.47</entry><entrymorerows="1">25.92</entry></column></row></column></row><row><column><entrymorerows="1">463</entry><entrymorerows="1">1425.65</entry><entrymorerows="1">22.34</entry><entrymorerows="1">513</entry><entrymorerows="1">1025.51</entry><entrymorerows="1">25.44</entry></column></row></column></row><row><column><entrymorerows="1">464</entry><entrymorerows="1">1451.71</entry><entrymorerows="1">29.19</entry><entrymorerows="1">514</entry><entrymorerows="1">3400.07</entry><entrymorerows="1">42.03</entry></column></row></column></row><row><column><entrymorerows="1">465</entry><entrymorerows="1">1576.66</entry><entrymorerows="1">26.5</entry><entrymorerows="1">515</entry><entrymorerows="1">1901.89</entry><entrymorerows="1">43.92</entry></column></row></column></row><row><column><entrymorerows="1">466</entry><entrymorerows="1">1651.85</entry><entrymorerows="1">40.6</entry><entrymorerows="1">516</entry><entrymorerows="1">1110.42</entry><entrymorerows="1">34.37</entry></column></row></column></row><row><column><entrymorerows="1">467</entry><entrymorerows="1">1876.94</entry><entrymorerows="1">22.29</entry><entrymorerows="1">517</entry><entrymorerows="1">1032.5</entry><entrymorerows="1">25.89</entry></column></row></column></row><row><column><entrymorerows="1">468</entry><entrymorerows="1">1911.12</entry><entrymorerows="1">24.98</entry><entrymorerows="1">518</entry><entrymorerows="1">1040.52</entry><entrymorerows="1">25.11</entry></column></row></column></row><row><column><entrymorerows="1">469</entry><entrymorerows="1">2064.01</entry><entrymorerows="1">21.95</entry><entrymorerows="1">519</entry><entrymorerows="1">1265.64</entry><entrymorerows="1">27.14</entry></column></row></column></row><row><column><entrymorerows="1">470</entry><entrymorerows="1">2150.04</entry><entrymorerows="1">27.76</entry><entrymorerows="1">520</entry><entrymorerows="1">1171.55</entry><entrymorerows="1">29.24</entry></column></row></column></row><row><column><entrymorerows="1">471</entry><entrymorerows="1">2751.59</entry><entrymorerows="1">29.16</entry><entrymorerows="1">521</entry><entrymorerows="1">1012.53</entry><entrymorerows="1">35.08</entry></column></row></column></row><row><column><entrymorerows="1">472</entry><entrymorerows="1">4289.94</entry><entrymorerows="1">28.69</entry><entrymorerows="1">522</entry><entrymorerows="1">1286.49</entry><entrymorerows="1">36.78</entry></column></row></column></row><row><column><entrymorerows="1">473</entry><entrymorerows="1">4306.05</entry><entrymorerows="1">28.78</entry><entrymorerows="1">523</entry><entrymorerows="1">2932.36</entry><entrymorerows="1">34.11</entry></column></row></column></row><row><column><entrymorerows="1">474</entry><entrymorerows="1">4800.18</entry><entrymorerows="1">23.83</entry><entrymorerows="1">524</entry><entrymorerows="1">1215.49</entry><entrymorerows="1">27.61</entry></column></row></column></row><row><column><entrymorerows="1">475</entry><entrymorerows="1">1111.32</entry><entrymorerows="1">35.47</entry><entrymorerows="1">525</entry><entrymorerows="1">1423.68</entry><entrymorerows="1">21.47</entry></column></row></column></row><row><column><entrymorerows="1">476</entry><entrymorerows="1">1181.49</entry><entrymorerows="1">36.79</entry><entrymorerows="1">526</entry><entrymorerows="1">1487.71</entry><entrymorerows="1">29.58</entry></column></row></column></row><row><column><entrymorerows="1">477</entry><entrymorerows="1">31688.38</entry><entrymorerows="1">24.69</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">478</entry><entrymorerows="1">1229.57</entry><entrymorerows="1">36.29</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">479</entry><entrymorerows="1">1579.78</entry><entrymorerows="1">29.83</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">480</entry><entrymorerows="1">1680.82</entry><entrymorerows="1">30.02</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">481</entry><entrymorerows="1">1725.66</entry><entrymorerows="1">38.3</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">482</entry><entrymorerows="1">5228.15</entry><entrymorerows="1">27.04</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">483</entry><entrymorerows="1">1769.78</entry><entrymorerows="1">28.25</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">484</entry><entrymorerows="1">1114.54</entry><entrymorerows="1">25.52</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">485</entry><entrymorerows="1">1390.5</entry><entrymorerows="1">37.05</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">486</entry><entrymorerows="1">2046.99</entry><entrymorerows="1">32.56</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">487</entry><entrymorerows="1">2899.33</entry><entrymorerows="1">49.62</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">488</entry><entrymorerows="1">1096.53</entry><entrymorerows="1">26.12</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">489</entry><entrymorerows="1">1257.49</entry><entrymorerows="1">34.26</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">490</entry><entrymorerows="1">868.45</entry><entrymorerows="1">23.35</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">491</entry><entrymorerows="1">1160.43</entry><entrymorerows="1">35.6</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">492</entry><entrymorerows="1">1539.8</entry><entrymorerows="1">40.36</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">493</entry><entrymorerows="1">3318.91</entry><entrymorerows="1">36.01</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">494</entry><entrymorerows="1">1084.48</entry><entrymorerows="1">25.31</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">495</entry><entrymorerows="1">1388.39</entry><entrymorerows="1">58.99</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">496</entry><entrymorerows="1">3129.86</entry><entrymorerows="1">35.93</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">497</entry><entrymorerows="1">1255.56</entry><entrymorerows="1">36.33</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">498</entry><entrymorerows="1">1383.69</entry><entrymorerows="1">39.02</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">499</entry><entrymorerows="1">1561.75</entry><entrymorerows="1">40.72</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></column></row><row><column><entrymorerows="1">500</entry><entrymorerows="1">3108.55</entry><entrymorerows="1">31.25</entry><entrymorerows="1"></entry><entrymorerows="1"></entry><entrymorerows="1"></entry></column></row></tbody></tgroup></column></row><table></tables>。2.根據(jù)權(quán)利要求l的方法，其特征在于利用下述參照值來評價對是否存在標志物1~106的確定<table>tableseeoriginaldocumentpage5</column></row><table>3.根據(jù)權(quán)利要求l的方法，其特征在于利用下述參照值來評價標:、物107~413的幅度<table>tableseeoriginaldocumentpage6</column></row><table><table>tableseeoriginaldocumentpage7</column></row><table>以及利用下述參照值來評價標志物414526的幅度:<table>tableseeoriginaldocumentpage8</column></row><table>4.根據(jù)權(quán)利要求1的方法，其中使用權(quán)利要求1中定義的至少兩種或至少三種或至少五種或六種或至少十種或所有多肽標志物。5.根據(jù)權(quán)利要求1至4中任一項的方法，其中所述來自受試者的樣品是尿樣品或血液樣品(血清或血漿樣品)。6.根據(jù)權(quán)利要求1至5中任一項的方法，其中毛細管電泳、HPLC、氣相離子色譜和/或質(zhì)譜用于檢測是否存在所述多肽標志物。7.根據(jù)權(quán)利要求1至6中任一項的方法，其中在測量所述多肽標志物分子質(zhì)量之前進行毛細管電泳。8.根據(jù)權(quán)利要求1至7中任一項的方法，其中質(zhì)譜用于檢測是否存在所述多肽標志物。9.選自標志物No.l-526的至少一種多肽標志物用于i貪斷血管病的用途，所述標志物的特征具有在于根據(jù)權(quán)利要求1的分子質(zhì)量值和遷移時間值。10.—種診斷血管病(VD)的方法，其包括以下步驟a)將樣品分成至少三個亞樣品，優(yōu)選至少十個亞樣品；b)分析至少兩個亞樣品以測定樣品中至少一種多肽標志物的存在與否或其幅度，其中所述多肽標志物選自標志物1~526,所述標志物的特征在于根據(jù)權(quán)利要求1的分子質(zhì)量和遷移時間(CE時間)。11.根據(jù)權(quán)利要求10的方法，其中測量至少十個亞樣品。12.根據(jù)權(quán)利要求1至11中至少一項的方法，其特征在于CE時間基于施加電壓為25kV、內(nèi)徑(ID)為50|um、長卯cm的玻璃毛細管，其中含20%乙腈、0.25M甲酸的水用作流動溶劑。13.—種標志物組合，其包含選自標志物1~526的至少十種標志物，所述標志物的特征在于具有根據(jù)權(quán)利要求1的分子質(zhì)量和遷移時間(CE時間)。全文摘要一種診斷血管病(VD)的方法。在所述方法中，測定樣品中是否存在至少一種多肽標志物，所述多肽標志物選自標志物1～526，所述標志物1～526的特征在于分子質(zhì)量值和遷移時間(CE時間)值。文檔編號G01N33/68GK101317095SQ200680044673公開日2008年12月3日申請日期2006年11月30日優(yōu)先權(quán)日2005年11月30日發(fā)明者哈拉爾德·米沙克申請人:馬賽奎斯診斷和治療有限公司