TWI599765B - 表面塗佈,使用表面塗佈捕捉、純化及釋放生物物質之方法與含有表面塗佈之新穎微流體晶片 - Google Patents
表面塗佈,使用表面塗佈捕捉、純化及釋放生物物質之方法與含有表面塗佈之新穎微流體晶片 Download PDFInfo
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- C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
- C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
- C07K16/30—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants from tumour cells
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- C—CHEMISTRY; METALLURGY
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- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
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- G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
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- G01N33/574—Immunoassay; Biospecific binding assay; Materials therefor for cancer
- G01N33/57484—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
- G01N33/57492—Immunoassay; Biospecific binding assay; Materials therefor for cancer involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites involving compounds localized on the membrane of tumor or cancer cells
Landscapes
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Description
本發明係有關於一種用於捕捉循環稀有細胞或循環腫瘤細胞之表面塗佈及包含此表面塗佈之新穎微流體晶片。
細胞脫落於循環中為惡性腫瘤之固有性質,且此特徵提供關於癌症患者之診斷、分級、治療反應及存活之重要資訊。舉例而言,Pantel等人發現血液中循環腫瘤細胞(Circulating Tumor Cell, CTC)的數目與癌症侵襲性以及療法功效有關。(Pantel, K.等人, 「Detection, clinical relevance and specific biological properties of disseminating tumor cells」, Nat Rev Cancer, 2008, 8(5):329-40)。
然而,在患有轉移性癌症之患者中,CTC為稀有細胞,每109個血細胞有一個CTC。此使得在技術上偵測及分離CTC具有難度(參見Kahn等人, Breast Cancer Res Treat 2004, 86:237-47)。因此,必須使用集中收集方法來有效偵測及分離CTC。
該集中收集方法之一實例為使用對CTC具有高特異性及敏感性之高度過度表現的細胞表面生物標記,諸如上皮細胞黏著分子(Epithelial Cell Adhesion Moledulce, EpCAM)。FDA批准的唯一CTC偵測平台Cellsearch System™(Veridex)利用抗EpCAM抗體塗佈之磁性奈米粒子捕捉及富集CTC,繼之以細胞角蛋白免疫染色(cytokeratin immunostaining)。另一市售CTC偵測系統AdnaTest(AdnaGen AG, Germany)採用藉由使用抗EpCAM及黏蛋白1(Mucin 1, MUC1)結合之磁性珠粒的類似免疫磁性方法。最近,開發基於抗EpCAM抗體塗佈之微流體晶片之「CTC晶片」用於CTC偵測及富集(Nagrath等人, Nature 2007, 450:1235-9)。然而,上述技術之缺陷為純CTC之低偵測率,此係歸因於血細胞與抗EpCAM抗體之非特異性結合。
為使CTC之偵測及分離達到最大程度,必須減少其他循環血細胞之非特異性結合。此舉可藉由使用生物惰性材料進行表面修飾來達成。舉例而言,Kaladhar等人觀察到顯著較少循環血細胞(例如血小板、白血球及紅血球)結合於經含有磷脂醯基-膽鹼、膽固醇及糖脂之各種脂質組合物之支撐單層修飾之固體基板上(Kaladhar等人, Langmuir 2004, 20: 11115-22及Kaladhar等人, J Biomed Mater Res A 2006, 79A:23-35)。
儘管偵測及分離CTC之技術在進步,仍需要特異性更高且更有效之方法來偵測、純化及釋放CTC及其他生物物質用於進一步培育及特性描述。
在一個態樣中,本發明係關於一種捕捉循環稀有細胞(CRC)之表面塗佈。此表面塗佈增加CRC,諸如CTC循環幹細胞(例如腫瘤幹細胞及骨髓幹細胞)、胎兒細胞、細菌、病毒、上皮細胞、內皮細胞或其類似物之捕捉效率且減少非特異性細胞之結合或蛋白質吸附。
表面塗佈包含1)減少非特異性血細胞結合及其他血液組分(諸如蛋白質)吸附之非沾黏材料組成;及2)捕捉CRC之生物活性組成。如圖1A所示,表面塗佈進一步包含連接至非沾黏材料組成及生物活性組成之連結分子。
在另一態樣中,本發明係關於一種捕捉及釋放生物物質之表面塗佈。此表面塗佈增加生物物質,諸如CTC循環幹細胞(例如腫瘤幹細胞、肝臟幹細胞及骨髓幹細胞)、胎兒細胞、細菌、病毒、上皮細胞、內皮細胞或其類似物之捕捉效率且增加非特異性細胞或蛋白質自該表面塗佈之移除或釋放。
表面塗佈包含1)自該表面塗佈釋放或移除非特異性血細胞及其他血液組分(諸如蛋白質)之可釋放組成;及2)捕捉生物物質之生物活性組成。表面塗佈進一步包含連接至可釋放組成及生物活性組成之連結分子。
本發明亦係關於一種微流裝置,其具有形成血液、體液或生物樣品之擾流以增加生物物質之捕捉率的特異性微結構設計。
本發明亦係關於一種製造表面塗佈之方法,其包含a)形成非沾黏材料或可釋放組成;及b)使連結分子與步驟a)之非沾黏材料/可釋放組成及生物活性組成連接,或c)使步驟a)之非沾黏材料/可釋放組成與生物活性組成連接。
本發明亦係關於自表面塗佈捕捉及釋放生物物質之方法。表面塗佈上之生物物質可藉由移除非特異性細胞或蛋白質來純化。所捕捉之生物物質可由氣泡、紫外線照射及其類似物釋放。
本發明亦係關於生物素標記之抗EpCam抗體EpAb4-1抗體捕捉CTC之用途。
本發明之實施例可參照隨附圖式進行描述。
本發明係關於一種表面塗佈,該表面塗佈有效捕捉循環稀有細胞(Circulating Rare Cell, CRC),諸如CTC循環幹細胞(例如腫瘤幹細胞及骨髓幹細胞)、胎兒細胞、細菌、病毒、上皮細胞、內皮細胞或其類似物。
在一個實施例中,用於捕捉CRC之表面塗佈包含1)防止結合非特異性細胞及吸附其他血液組分(諸如蛋白質)之非沾黏材料組成;及2)捕捉循環稀有細胞之生物活性組成。非沾黏材料組成及生物活性組成藉由該等非沾黏材料組成及生物活性組成中存在的離散官能基或部分接合。兩種組合物之間的鍵結通常藉由相互作用形成,該相互作用包含靜電相互作用、親水性-親水性相互作用、極性-極性相互作用、互補DNA結合、磁力或其組合。
在一組實施例中,互補DNA片段用於結合非沾黏材料組成及生物活性組成。片段連接於各組合物且可在其長度上部分或完全互補。DNA之合適長度通常為至少15、20、25、35、50、100或100個以上鹼基長。本發明中所用之DNA之一實例為DNA鉗。(參見B Yurke等人, A DNA-fuelled molecular machine made of DNA. Nature 2000, 406:605-608)。
在另一組實施例中,表面塗佈包含1)非沾黏材料組成;2)生物活性組成;及3)連結分子,其使非沾黏材料組成接合於生物活性組成。參見圖1A。
本發明亦係關於一種表面塗佈,其有效捕捉生物物質,諸如CTC循環幹細胞(例如腫瘤幹細胞、肝臟幹細胞及骨髓幹細胞)、胎兒細胞、細菌、病毒、上皮細胞、內皮細胞或其類似物;藉由經緩衝液沖洗釋放或移除非特異性細胞及其他血清組分(例如蛋白質)來純化該表面塗佈表面上之生物物質;及自該表面塗佈釋放所捕捉之生物物質。
用於捕捉及純化生物物質之表面塗佈包含1)經由緩衝液沖洗釋放非特異性血細胞及其他血液組分(諸如蛋白質)之可釋放組成;及2)捕捉生物物質之生物活性組成。可釋放組成及生物活性組成藉由該等可釋放組成及生物活性組成中存在的離散官能基或部分接合。兩種組合物之間的鍵結通常藉由相互作用形成,該相互作用包含靜電相互作用、親水性-親水性相互作用、極性-極性相互作用、互補DNA結合、磁力或其組合。
在一個實施例中,表面塗佈進一步包含連接至可釋放組成及生物活性組成之連結分子。
如下文更詳細地闡明,表面塗佈可併入以下組態中:細胞培養皿、微流體通道、微流體晶片、過濾器、毛細管、試管、珠粒、奈米粒子或其類似物,內徑在約50 μm至約1000 μm之範圍內。
非沾黏材料及可釋放組成
「非沾黏材料」組成(參見圖1A)減少非特異性細胞結合及血清蛋白吸附。
「可釋放」組合物包含亦充當「潤滑」表面以使得自表面塗佈移除或釋放非特異性細胞或血液組分僅需要低流動剪切應力同時保持生物物質完整的非沾黏材料組成。
非沾黏材料組成係選自由以下組成之群:支撐脂質層,諸如脂質體、支撐脂質雙層(SLB)或脂質多層;多肽;聚電解質多層(PEM);聚乙烯醇;如圖2A所示之聚乙二醇(PEG);水凝膠聚合物;細胞外基質蛋白;碳水化合物;聚合物刷;兩性離子材料,諸如如圖2D所示之聚(羧基甜菜鹼)(pCB)、如圖2E所示之聚(磺基甜菜鹼)(pSB)及pDMAEMA;小有機化合物;及形成單層或多層之上述材料之組合。
對於其中非沾黏材料組成包含支撐脂質雙層(SLB)之彼等實施例,SLB通常包含諸如以下之脂質:例如,如圖2B所示之1,2-二油醯基-sn-甘油-3-磷酸乙醇胺-N-(cap生物素基)(鈉鹽)(b-PE)及1-棕櫚醯基-2-油醯基-sn-甘油-3-磷酸膽鹼(POPC)。SLB之蛋白質抗性可由在寬pH值範圍中存在中性及兩性離子磷脂醯膽鹼頭基以及親水性脂質頭基與本體溶液之間形成水性薄膜來解釋(參見Johnson等人, Biophys J 1991, 59:289-94)。
在另一組實施例中,非沾黏材料組成包含PEG,較佳具有約100至約100,000之分子量且顯示非沾黏材料性的PEG。
在又一組實施例中,非沾黏材料組成包含聚電解質多層(PEM)或聚合物刷。適用於本發明之PEM之實例包括(但不限於)聚-L-離胺酸/聚-L-麩胺酸(PLL/PLGA)、聚-L-離胺酸/聚-L-天冬胺酸或類似抗衡離子聚電解質。聚合物刷包含如圖2C所示之(氯化[2-(丙烯醯基氧基)乙基]三甲基銨,TMA)/(丙烯酸2-羧乙酯,CAA)共聚物。非沾黏材料層通常具有幾奈米直至幾百微米之厚度。
非沾黏材料組成包含能夠共價、非共價或共價及非共價組合連接之官能基,該等官能基直接連接於生物活性組成中存在的官能基或直接連接於作為鍵結組合物之一部分的官能基。
在一些實施例中,非沾黏材料組成之官能基(在共價連接前)係選自:羥基、胺基、羧酸或酯基、硫酯基、醛基、環氧基或環氧乙烷基、肼基及硫醇基,該等官能基經選擇以與連結分子或生物活性組成中存在的官能基反應。在其他實施例中,非沾黏材料組成之官能基(在非共價連接前)作為結合對第一成員係選自由生物素、抗生物素蛋白、抗生蛋白鏈菌素、DNA、RNA、配位體、受體、抗原、抗體及正負電荷組成之使用特異性結合識別之群,經選擇結合對第一成員結合於連結分子或生物活性組成中存在的結合對第二成員。
連結分子接合非沾黏材料/可釋放組成與生物活性組成且包含官能基,該等官能基能夠共價、非共價或共價及非共價組合直接連接於非沾黏材料/可釋放組成中存在之官能基且直接連接於作為生物活性組成之一部分的官能基。
在一些實施例中,連結分子包含選自以下之官能基(在共價連接前):羥基、胺基、羧酸或酯基、硫酯基、醛基、環氧基或環氧乙烷基、肼基及硫醇基,該等官能基經選擇與非沾黏材料組成或生物活性組成中存在的官能基反應。
在其他實施例中,連結分子包含作為結合對第一成員之官能基(在非共價連接前),該等官能基選自由生物素、抗生物素蛋白、抗生蛋白鏈菌素、DNA、RNA、配位體、受體、抗原、抗體及正負電荷組成之使用特異性結合識別之群,經選擇結合對第一成員結合於非沾黏材料/可釋放組成或生物活性組成中存在之結合對第二成員。
連結分子上之官能基亦可為選自以下之可切割官能基:可藉由紫外線照射裂解之光敏官能基、可藉由電脈衝機制裂解之電敏官能基、可藉由磁力消除裂解之磁性材料、可藉由破壞靜電相互作用裂解之聚電解質材料、可藉由雜交裂解之DNA及其類似物。
生物活性組成:
生物活性組成接合於連結分子或非沾黏材料組成,且包含用於選擇性偵測生物物質或CRC之結合部分。
生物活性組成包含官能基,該等官能基能夠共價、非共價或共價及非共價組合直接連接於非沾黏材料層中存在的官能基或直接連接於作為連結分子之一部分的官能基。
在一些實施例中,生物活性組成之官能基(在共價連接前)係選自:羥基、胺基、羧酸或酯基、硫酯基、醛基、環氧基或環氧乙烷基、肼基及硫醇基,該等官能基經選擇與非沾黏材料組成或連結分子中存在的官能基反應。在其他實施例中,生物活性組成之官能基(在非共價連接前)選自由生物素、抗生物素蛋白、抗生蛋白鏈菌素、DNA、RNA、配位體、受體、抗原-抗體及正負電荷組成之使用特異性結合識別之群,經選擇之官能基結合於非沾黏材料/可釋放組成或連結分子上存在之結合對第二成員。
生物活性組成之結合部分與生物物質經由分子識別、化學親和力或幾何/形狀識別具有特異性親和力。用於探測生物物質之結合部分的實例包括(但不限於):合成聚合物、分子印記聚合物、細胞外基質蛋白、結合受體、抗體、DNA、RNA、抗原或對生物物質呈現高親和力之任何其他表面標記。較佳抗體為抗EpCAM膜蛋白抗體(可購自許多來源,包括R&D Systems, MN, USA),其提供對於CTC之高特異性,這是因為EpCAM常常過度表現於肺惡性腫瘤、結腸直腸惡性腫瘤、乳房惡性腫瘤、前列腺惡性腫瘤、頭頸部惡性腫瘤及肝臟惡性腫瘤中,但血細胞中不存在。另一較佳抗體為抗HER2,其對於CTC具有高特異性但不存在於血細胞中。
在一個實施例中,抗EpCAM膜蛋白抗體為EpAb4-1抗體,包含表1中展示之SEQ ID No:1重鏈序列及SEQ ID NO: 2輕鏈序列。 表1. EpAb4-1抗體之VH
及VL
結構域之胺基酸序列
展示VH結構域及VL結構域兩者之互補決定區1-3(CDR1-3)、構架區1-4(FW1-4)。V結構域家族由VBASE2資料庫(www.vbase2.org)比對。
生物活性組成可具有多種厚度,選擇生物活性組成之厚度不影響表面塗佈之功能或效能。
在一個實施例中,非沾黏材料組成及生物活性組成之結合連結分子或催化劑為生物素/抗生物素蛋白或其衍生物。在另一實施例中,非沾黏材料組成及生物活性組成之結合連結分子或催化劑為EDC/NHS。在另一較佳實施例中,非沾黏材料組成及生物活性組成之結合連結分子或催化劑為磺基-SMCC。圖3圖解說明此等實施例之化學反應。
固體基板:
在一些實施例中,如圖4A所示,表面塗佈在沒有表面連結分子的情況下連接於固體基板。非沾黏材料/可釋放組成經由以下相互作用中之一者連接於固體基板:共價鍵(對於PEG非沾黏材料組成)、氫鍵、靜電相互作用、親水性-親水性相互作用(對於SLB非沾黏材料/可釋放組成)、極性-極性相互作用、互補DNA結合、磁力或其類似物。
在其他實施例中,如圖4D所示,表面塗佈使用表面連結分子連接於固體基板。用於本發明之固體基板的實例包括(但不限於):金屬、塑膠、玻璃、矽晶圓、羥基化聚(甲基丙烯酸甲酯)(PMMA)及其組合。固體基板之形狀包括(但不限於):平面、圓形及具有微米結構或奈米結構之不規則形狀,諸如奈米粒子、奈米線及其組合。
表面連結分子包含官能基,該等官能基能夠共價、非共價、或共價及非共價組合直接連接於非沾黏材料/可釋放組成中存在之官能基及直接連接於作為固體基板之一部分的官能基。用於使表面塗佈結合於玻璃基板之表面連結分子的實例包括(但不限於)矽烷、胺基丙基三乙氧基矽烷、胺基丙基三甲氧基矽烷、矽烷-PEG-NH2、矽烷-PEG-N3(PEG分子量為約1,000至約30,000道爾頓)及矽烷-PEG生物素。
在一組實施例中,表面連結分子包含選自以下之可切割官能基:可藉由紫外線照射裂解之光敏官能基、可藉由電脈衝機制裂解之電敏官能基、藉磁力消除釋放非沾黏材料組成之鐵或磁性材料、可藉由破壞靜電相互作用裂解之聚電解質材料、可藉由雜交裂解之DNA或及其類似物。
在一個實施例中,非沾黏材料組成包含矽烷-官能化PEG且固體基板較佳選自由矽、玻璃、羥基化聚(甲基丙烯酸甲酯)(PMMA)、氧化鋁、TiO2及其類似物組成之群。在另一實施例中,非沾黏材料組成包含硫醇-官能化化合物且固體基板較佳選自由Au、Ag、Pt及其類似物組成之群。
製造表面塗佈之方法:
圖5A及5B展示形成表面塗佈之步驟:
1.形成具有合適官能基(生物素)之非沾黏材料/可釋放組成(例如SLB或PEG);
2.使連結分子上之官能基(抗生蛋白鏈菌素)連接於非沾黏材料/可釋放組成上之官能基(生物素);
3.形成生物活性組成,且使生物活性組成上之官能基(生物素)連接於連結分子上之官能基(抗生蛋白鏈菌素)。(以下稱為「段落A」)
無連結分子之表面塗佈可藉由以下步驟形成:
1.形成具有合適官能基(例如N-戊二醯基磷脂醯乙醇胺或NGPE之羧基)之非沾黏材料/可釋放組成;
2.形成且使生物活性組成上之官能基(一級胺)連接於步驟1中非沾黏材料/可釋放組成上之官能基(NGPE之羧基)。(以下稱為「段落B」)
段落A與段落B中之步驟可顛倒。
微流體晶片:
如圖6A所示,微流體晶片包含第一固體基板1(例如PMMA)及第二固體基板2(例如玻璃),其中該第一及第二固體基板1、2使用黏著構件3或其他構件黏著在一起。
參照圖6B,一個或兩個固體基板之表面可雕刻有微結構4。在一組實施例中,微結構4以線性方式排列。在另一組實施例中,微結構4以人字形方式排列。切除圖6B中黏著構件3上之陰影區以容納固體基板1表面上之微結構4。藉由將第一固體基板1及第二固體基板2與黏著構件3黏著在一起形成密封通道5。密封通道5之高度由黏著構件3之厚度決定。
一旦形成微流體晶片,表面塗佈則可連接於一或兩個固體基板。在一組實施例中,表面塗佈使用表面連結分子連接於固體基板。在另一組實施例中,表面塗佈經由以下相互作用中之一者連接於固體基板:共價鍵(對於PEG非沾黏材料組成)、氫鍵、靜電相互作用、親水性-親水性相互作用(對於SLB非沾黏材料/可釋放組成)、極性-極性相互作用、互補DNA結合、磁力或其類似物。
參照圖6C,固體基板1上之微結構4垂直於流動方向且當血液、體液或生物樣品通過微流體晶片之密封通道5時,形成血液、體液或生物樣品之混沌流(chaotic flow)或擾流。擾流增加生物物質-表面塗佈接觸。
兩種因素支配微流體晶片之捕捉效率:
(1)血液、體液或生物樣品之線性速度,其決定生物物質與表面塗佈之接觸時間。在一個較佳實施例中,線性速度為約0.1 mm/s至1 mm/s。在一個更佳實施例中,圖7F中設計E之線性速度為約0.42 mm/s或0.5 ml/h。
(2)由固體基板上之微結構4所引起的血液、體液或生物樣品之流動擾動。流動擾動增加生物物質與表面塗佈之間的接觸。(下文稱為「段落C」)
圖7A展示固體基板上微結構4之不同設計。設計F中之微結構以人字形圖案排列,而設計A-E及H中之微結構以線性圖案排列。微結構4之尺寸如下:O-D及G之長度為約50 mm且E-F之長度為約120 mm,高度為約30 μm,O及A之寬度為約1.5 mm,B之寬度為約3.0 mm且C-G之寬度為約5.5 mm。密封通道5之高度隨黏著構件3之厚度而變化,較佳為約30-90 μm,更佳為約60 μm。
圖7B-7H展示圖7A中設計A-G之細節。圖7H中之設計G為具有以下尺寸之較佳圖案:微結構寬度(W)為約150 μm,微結構長度(L)為約1000 μm,兩列微結構之間的距離(Sr)為約250 μm,兩個相鄰微結構之間的距離(Sc)為約350 μm,微結構高度(D)為約30 μm且密封通道5之高度(H)為約60 μm。
不同設計之生物物質捕捉效率展示於圖7I及圖7J中。捕捉率定義為(所捕捉之生物物質/測試樣品中之初始生物物質)×100%。通道O不具有微結構且具有最低生物物質捕捉率,DMEM樣品及血液樣品之最低生物物質捕捉率分別為27%及1%。設計E對於摻料於DMEM中之HCT116癌細胞具有80%捕捉率,且對於摻料於血液樣品中之HCT116癌細胞具有30%捕捉率。設計F具有最佳捕捉率,捕捉平均超過70%摻料於血液樣品中之HCT116癌細胞(參見圖7J)。
流動純化:
表面塗佈上之生物物質可藉由移除非沾黏材料/可釋放組成表面上之非特異性細胞及其他血液組分而進一步純化。非沾黏材料/可釋放組成對於非特異性細胞及其他血液組分具有低親和力。因此,用約0.8 dyne/cm2
至約50 dyne/cm2
之低流量緩衝溶液沖洗表面塗佈足以移除非沾黏材料/可釋放組成上之非特異性細胞及其他血液組分,而生物物質保留於表面塗佈上。
在一個較佳實施例中,緩衝液沖洗剪切力為約2.5 dyne/cm2
至約10 dyne/cm2
。圖8展示當緩衝液流之剪切應力為約3.3 dyne/cm2
時,移除80%之非特異性細胞(亦即白血球),同時無生物物質(亦即HCT 116癌細胞)自表面塗佈移除。當緩衝液流之剪切應力增至8 dyne/cm2
時,幾乎移除所有非特異性細胞,同時無生物物質自表面塗佈移除。
釋放生物物質:
在藉由流動純化移除大部分非特異性細胞及血液組分後,可自表面塗佈釋放生物物質。
若非沾黏材料/可釋放組成包含脂質或脂質混合物,則所捕捉之生物物質可藉由引入氣泡溶液或油相釋放。如圖9中所示,表面塗佈包含非沾黏材料組成A(脂質雙層)及生物活性組成B(抗體)且結合於固體基板S。生物物質CTC結合於生物活性組成B,而其他細胞受非沾黏材料組成A排斥。隨著氣泡接近脂質雙層,脂質雙層之疏水性尾因其與氣泡內部亦為疏水性之空氣的高親和力而上下顛倒。此舉破壞脂質雙層表面上之親水性-親水性相互作用且使氣泡「抬起」脂質雙層頂層以及生物活性組成上結合之CTC。
若非沾黏材料組成包含除脂質或脂質混合物以外之組合物,則所捕捉之生物物質可藉由破壞連結分子或表面連結分子上之可切割官能基而釋放。此釋放機制於圖10A及10B中說明。圖10A展示固體基板上之表面塗佈,其中該表面塗佈包含生物活性組成B、具有可切割官能基C之連結分子及非沾黏材料組成A。表面塗佈藉由表面連結分子1連接於固體基板S(例如玻璃)。圖10B展示自圖10A之表面塗佈釋放生物物質(例如CTC)。生物物質結合於生物活性組成B,而其他細胞受非沾黏材料組成A排斥。用365 nm紫外光照射表面塗佈,由此破壞連結分子C上之可切割官能基,且釋放生物物質以供後續分析但維持其生存力。
生物物質亦可藉由其他機制釋放。在一組實施例中,連結分子或表面連結分子包含電敏可切割官能基,且藉由電脈衝機制釋放生物物質。在另一組實施例中,連結分子或表面連結分子包含磁性材料作為可切割官能基,且磁場或磁力消除時釋放生物物質。在又一組實施例中,連結分子或表面連結分子包含PEM作為可切割官能基,且藉由改變層間靜電相互作用釋放生物物質。在又一組實施例中,連結分子或表面連結分子包含DNA片段作為可切割官能基,且藉由DNA雜交釋放生物物質。
實例:
以下實例進一步說明本發明。此等實例僅意欲說明本發明且不應理解為限制本發明。
實例1:製備雙層表面塗佈
製備非沾黏材料組成:
藉由以下步驟製備支撐脂質雙層(SLB):
(1)將POPC及b-PE(可購自Avanti Polar Lipids, USA)溶解於氯仿中且最終脂質濃度為5 mg/ml。在緩慢氮氣流下旋風乾燥POPC/b-PE溶液,形成均勻的POPC/b-PE薄膜。在真空室中進一步乾燥POPC/b-PE薄膜隔夜以移除殘餘氯仿。
(2)將步驟(1)之POPC/生物素-PE膜分散於含有10 mM磷酸鹽緩衝鹽水、150 mM氯化鈉水溶液及0.02%(w/v)疊氮化鈉(NaN3,可購自Sigma-Aldrich, USA)之磷酸鹽緩衝液(其中pH值調節至7.2)中且與其混合。在150 psi下在室溫下,經由100 nm隨後50 nm Nuclepore®徑跡蝕刻聚碳酸酯膜片(Whatman Schleicher & Schuell, Germany)過濾混合溶液至少10次。
(3)使步驟(2)中經過濾之溶液流經LIPEX™擠壓機(Northern Lipids, Inc. Canada),生成單層微脂粒之均勻群體。由動態雷射光散射偵測器(Zetasizer Nano ZS, Malvern Instruments, Germany)測定之POPC/生物素-PE微脂粒之尺寸為約65±3 nm。
製備生物活性組成:
由以下步驟製備生物素標記之EpCAM抗體:
(1)藉由Chen等人(Clin Vaccine Immunol 2007;14:404-11)所述之方法製造抗EpCAM單株抗體(OC98-1或EpAb4-1)。
(2)將步驟(1)中之抗體溶解於含有10 mM PBS及150 mM NaCl之緩衝溶液(pH值為約7.2)中。由Nanodrop 1000分光光度計(Thermo Scientific, USA)所測定,抗體緩衝溶液濃度為約0.65 mg/mL。
(3)使步驟(2)之抗體溶液與10 mM磺基NHS-LC-生物素混合(莫耳比為1比10)且在室溫下溶解於Milli-Q水(Milli-Q RO系統, USA)中30分鐘。過量生物素藉由在4℃下於磷酸鹽緩衝鹽水中透析24小時(每12小時更換緩衝液)來移除。
(4)使用生物素定量套組(Pierce, USA)藉由HABA檢定所測定,生物素標記之抗EpCAM抗體(bOC98-1或bEpAb4-1)中生物素與抗體之比率為1.5比1。
或者,可使用R and D Systems(Minneapolis, MN)市售之生物素標記之山羊抗人類抗EpCAM抗體。
製備本發明之固體基板:
玻璃基板(諸如Deckglaser, Germany之顯微鏡蓋玻片)用10% DECON 90(Decon Laboratories L imited, England)清潔,用Milli-Q水沖洗,在氮氣下乾燥,且暴露於電漿清潔器(Harrick Plasma, Ithaca, NY, U.S.A.)中100 mtorr下之氧電漿10分鐘。在每次使用之前,用乙醇沖洗玻璃基板且在氮氣下乾燥。
在120℃下,用強清潔性溶液(70%硫酸及30%過氧化氫(v/v))清潔基於二氧化矽之固體基板(例如矽晶圓或玻璃蓋玻片),隨後用蒸餾水洗滌且用丙酮沖洗。在氮氣流下乾燥固體基板且用電漿清潔器處理。
對於汽相矽烷化反應,將清潔的二氧化矽基板及含有150 μL 3-(胺基丙基)-三乙氧基矽烷(Sigma, USA)之皮氏培養皿(Petridish)置於約0.3托之減壓乾燥器(Wheaton dry-seal desiccator, 100 nm)中16小時。由丙酮清潔基板且在氮氣流下乾燥。
在固體基板上構建SLB表面塗佈:
添加0.25 mg/mL段落C之POPC/b-PE微脂粒溶液至經清潔之固體基板上,形成經SLB塗佈之固體基板。接著用含有10 mM PBS及150 mM NaCl(pH值= 7.2)之磷酸鹽緩衝液澈底沖洗此固體基板,以移除過量POPC/b-PE微脂粒。生物素為SLB中與連結分子中之官能基(抗生蛋白鏈菌素)結合之官能基。
添加0.1 mg/mL抗生蛋白鏈菌素(Streptavidin, SA)溶液(可購自Pierce Biotechnology, Rockford, IL, USA)至經SLB塗佈之固體基板上且培育1小時,接著用PBS緩衝液沖洗以移除過量SA。
添加約0.05 mg/mL b-抗EpCAM溶液至經SA-SLB塗佈之固體基板上,形成本發明之表面塗佈。
在固體基板上構建PEG表面塗佈:
添加生物素標記之PEG矽烷溶液(Si-bPEG)至清潔的玻璃基板上且培育1小時,以便在玻璃基板上形成Si-bPEG非沾黏材料組成,隨後乙醇沖洗以移除過量Si-bPEG。矽烷為表面連結分子且生物素為與連結分子中之官能基(SA)結合的官能基。
添加0.1 mg/mL SA溶液至經Si-bPEG塗佈之固體基板上且培育1小時,隨後PBS緩衝液沖洗以移除過量SA。
添加0.05 mg/mL b-抗EpCAM溶液且與SA-Si-bPEG表面塗佈結合,隨後PBS緩衝液沖洗以移除過量b-抗EpCAM。
在固體基板上構建PEM表面塗佈:
分批且在如下固定條件下進行PEM膜之物理沈積:起初,將所有多肽溶解於10 mM Tris-HCl緩衝液及0.15 M NaCl(pH 7.4)中。接著在室溫下,使固體基板浸沒於PLL(MW 15000-30000;Sigma, St Louis, MO)溶液(1 mg/mL)中10分鐘,隨後用1 mL Tris-HCl緩衝液沖洗1分鐘。隨後,為偶合PLGA,將經PLL塗佈之載玻片浸沒於PLGA溶液(MW 3000-15000,Sigma, St Louis, MO,1 mg/mL)中10分鐘,隨後用1 mL Tris-HCl緩衝液沖洗1分鐘。最後,用新鮮PBS清潔基板以移除未偶合之多肽。結果c-(PLL/PLGA)i,其中i表示藉由重複上述步驟生成之聚電解質對的數目:i)0.5僅指c-PLL,i)1指c-(PLL/PLGA)1,及其類似物。
SLB表面塗佈之QCM-D表徵:
藉由耗散型石英晶體微量天平(QCM-D)監測表面塗佈之構建。圖11中之QCM-D反應展示在經SiO2預處理之石英晶體上的表面塗佈構建。首先,在點(I)處,將0.25 mg/mL POPC/b-PE微脂粒混合物(於磷酸鹽緩衝液中)施配於QCM室中。歸一化頻率變化F及耗散位移D分別為26.0±0.7 Hz及0.19±0.03×10-6,其為高度均勻的脂質雙層之特徵。在兩次緩衝液洗滌(表示為*)後,在點II處施配0.1 mg/mL SA溶液。SA結合在F = 52.8±5.4 Hz及D = 3.84±0.54×10-6時飽和。在點(III)處,將0.025 mg/mL OC98-1抗體溶液施配於QCM室中且不存在頻率或耗散變化。由此可見在OC98-1抗體與SA-脂質雙層表面之間不存在相互作用。相比之下,在點(IV)處添加生物素標記之抗體溶液(bOC98-1或bEpAb4-1)產生頻率及耗散變化,平衡位移F = 39.4±6.8 Hz及D = 1.63±0.28×10-6。此種情況證明生物素標記之抗體結合於SA-脂質雙層表面。
使用QCM-D檢查表面塗佈上SLB非沾黏材料組成之特徵(圖12)。添加牛血清白蛋白(BSA,可購自Sigma-Aldrich, USA)至表面塗佈上且頻率及耗散驟然變化,平衡位移F = 6.9 Hz及D = 3.35×10-6。此情況表明即刻BSA吸附。三次緩衝液沖洗(*)引起頻率增加及耗散降低,飽和位移F = 6.1 Hz及D = 3.16×10-6。此情況表明所吸附之BSA可易於自表面塗佈移除,且因此表明BSA與SLB之間非常弱的相互作用。
實例2:製備微流體晶片
微流體晶片可藉由以下步驟製備:
1.使用商用CO2雷射雕刻器(Helix 24, Epilog, USA)雕刻微槽,以便在PMMA基板上形成微結構。
2.用MeOH、清潔劑及水,隨後10分鐘音波處理清潔PMMA基板、玻璃基板及螺帽。螺帽及固體基板由氮氣乾燥且在60℃下烘烤10分鐘。
3.藉由氯仿處理,使PMMA基板與螺帽黏結在一起。
4.使用黏著構件(例如3M, USA之3M雙面膠帶)將PMMA基板與玻璃載玻片接合在一起。
實例3:CTC結合於抗EpCAM官能化SLB表面塗佈
使用八個血液樣品測定實例2之微流體晶片中抗EpCAM官能化SLB表面塗佈之CTC捕捉率。各血液樣品含有2 ml IV期結腸癌患者血液,且樣品由注射泵控制以0.5毫升/小時引入微流體晶片之密封通道。隨後,用0.5 ml PBS緩衝液在1毫升/小時之流動速率下沖洗微流體晶片中之密封通道,接著原位免疫染色。
此8種樣品每毫升血液所捕捉之CTC數目為26、34、36、39、47、67、79及99。25%之血液樣品具有每毫升測試樣品79或79以上之CTC計數,且中值CTC計數為每毫升測試樣品43。在緩衝液沖洗後,非特異性細胞與蛋白質之結合極少。
作為比較,FDA批准之Veridex CellSearch之CTC計數如下:25%之樣品具有每7.5 ml測試樣品3或3個以上CTC且中值CTC計數為0。
抗EpCAM官能化SLB表面與150 μL含有HCT116癌細胞之人類血液(HCT116癌細胞密度為每100 μL血液約10至100個)一起培育,接著緩衝液沖洗移除非特異性細胞。圖13展示緩衝液沖洗之前及之後之表面塗佈。在緩衝液沖洗之前,表面塗佈由非特異性細胞(左上)及四個HCT116癌細胞(左下)覆蓋。在緩衝液沖洗後,移除幾乎所有非特異性細胞(右上),但四個HCT116癌細胞(右下)保留於表面塗佈上。
結果顯示本發明之表面塗佈有效捕捉CTC及釋放非特異性細胞。
實例4:各種表面條件之捕捉效率及非沾黏材料性質的比較
HCT116癌細胞(生物物質)之捕捉率及6種不同表面條件之非沾黏材料性質展示於圖14A。
結果顯示本發明之表面塗佈(脂質/SA/b-抗EpCAM及PEG(15 mM)/SA/b-抗EpCAM)在捕捉生物物質方面更有效。與沒有非沾黏材料組成之表面塗佈(僅玻璃)相比,本發明之表面塗佈上非特異性細胞(白血球或WBC)之結合較少。
圖14B展示以下表面之非特異性血細胞結合:(A)僅玻璃;(B)生物素標記之SLB(b-SLB),(C)抗生蛋白鏈菌素結合之bSLB,及(D)OC98-1結合之bSLB。此等表面與來自健康供體之稀釋的人類血液(含1 μL血液之100 μL PBS緩衝液)一起培育4小時,接著PBS緩衝液沖洗。影像(E)至(H)為對應於(A)至(D)之表面塗佈的沖洗後影像。結果顯示與無可釋放組成之表面塗佈(亦即僅玻璃)相比,在緩衝液沖洗後,具有可釋放組成(亦即SLB)之表面塗佈上的非特異性血細胞較少。
實例5:藉由流動進行純化
差異流動剪切力可基於此等細胞與非沾黏材料組成之親和力選擇性「清除(flush)」非特異性細胞,而生物物質保留於表面塗佈上。
在此研究中,表面塗佈包含SLB、連結分子及作為生物活性組成之纖維結合蛋白。圖15A展示纖維母細胞3T3(綠色)及結腸癌細胞株HCT116(紅色)在表面塗佈上培育4小時。用具有3 dyne/cm2
之剪切應力的緩衝溶液沖洗表面塗佈。
如圖15B所示,在緩衝液沖洗5分鐘內,自表面塗佈清除掉HCT 116細胞(紅色)。如圖15C所示,纖維母細胞3T3細胞(綠色)因其對纖維結合蛋白之高親和力而在30分鐘緩衝液沖洗後保留於表面塗佈上。
結果顯示約3 dyne/cm2
之剪切應力足以自可釋放組成移除非特異性細胞。
圖16概述HCT116及NIH-3T3細胞群體(非特異性細胞)之各別剪切應力及沖洗時間。為自表面塗佈之可釋放組成移除HCT116細胞,剪切應力為約3 dyne/cm2
至約4.5 dyne/cm2
。為自表面塗佈之可釋放組成移除NIH-3T3細胞,剪切應力為約8.5 dyne/cm2
至約12 dyne/cm2
(N/N0為使用各種剪切應力仍連接於表面塗佈之細胞百分比。N為最終細胞數目且N0為初始細胞數目。)
實例6:自表面塗佈釋放CTC
實例3中表面塗佈上所捕捉之HCT116癌細胞藉由引入氣泡釋放。圖17展示紅圈中之HCT116細胞在引入氣泡3秒內自表面塗佈移除。
實例7:培養自表面塗佈釋放之CTC
所捕捉之CTC與5 mM EDTA一起在37℃下培育5至10分鐘,且藉由流動培養基釋放至微流體晶片之密封通道中。由此程序總共釋放18個colo205細胞。將所釋放之colo205細胞與含血清之培養基及抗生素(青黴素+鏈黴素+慶大黴素)一起置於48孔組織培養聚苯乙烯板中進行培育。
圖18展示18個colo205細胞分別在第1天(小圖A)、第10天(小圖B)及第14天(小圖C)之一部分。此研究證明所釋放之colo205細胞在後續細胞培養中保留其生存力。
實例8:經由CTC過濾裝置捕捉CTC
可用本發明之表面塗佈塗佈任何膜片、試管、毛細管、珠粒、奈米粒子或通道。圖19圖解說明一種過濾裝置,其中該過濾器經本發明之表面塗佈塗佈。該過濾器可容納高容量之血流且捕捉用於診斷或治療目的之生物物質。為獲取患者血液或體液,可將導管插入患者靜脈或瘺管中且患者血液流經CTC過濾裝置,其中過濾器上之表面塗佈捕捉CTC。經過濾之血液流回患者。
實例9:經由生物素標記之EpAb4-1抗體捕捉CTC
使用HCT116(結腸直腸)CTC及SAS(舌)CTC檢查生物素標記之OC9801抗體、生物素標記之EpAb4-1抗體及生物素標記之EpCam抗體(可購自R&D system, USA)之結合特異性。
CTC摻雜於緩衝溶液中(每毫升約105個CTC)。將摻雜CTC之緩衝溶液與以下生物活性組成一起引入表面塗佈:生物素標記之OC9801抗體、生物素標記之EpAb4-1抗體、生物素標記之EpCam抗體及IgG抗體。
藉由比色法,量測在490 nm下之吸收光密度來測定抗體之CTC結合特異性。圖20展示生物素標記之EpAb4-1有效捕捉HCT116 CTC及SAS CTC。
1‧‧‧第一固體基板
2‧‧‧第二固體基板
3‧‧‧黏著構件
4‧‧‧微結構
5‧‧‧密封通道
2‧‧‧第二固體基板
3‧‧‧黏著構件
4‧‧‧微結構
5‧‧‧密封通道
圖1A圖解說明包含非沾黏材料組成、連結分子及生物活性組成之表面塗佈之一實施例。 圖1B圖解說明循環腫瘤細胞與圖1A之表面塗佈的結合。 圖2A至圖2E說明非沾黏材料材料之實例之化學結構。 圖3說明在非沾黏材料組成上之官能基與生物活性組成之間結合的化學反應。 圖4A圖解說明表面塗佈與固體基板在沒有表面連結分子之情況下的連接。 圖4B及圖4C圖解說明具有可切割官能基之連結分子。 圖4D圖解說明表面塗佈與固體基板使用表面連結分子的連接。 圖5A及圖5B圖解說明在固體基板上形成表面塗佈。 圖6A及圖6B圖解說明微流體晶片之組件。 圖6C圖解說明自生物樣品捕捉CTC之微流體晶片總成。 圖7A至圖7H圖解說明固體基板上之微結構設計。 圖7I及圖7J說明各種微結構設計分別在DMEM溶液及血液中之捕捉效率。 圖8說明緩衝溶液釋放非特異性細胞及純化所捕捉之生物物質的剪切應力。 圖9圖解說明由氣泡法釋放生物物質。 圖10A圖解說明在固體基板上具有可分割連結分子之表面塗佈。 圖10B圖解說明自圖10A之表面塗佈釋放生物物質。 圖11說明表面塗佈構建之QCM-D反應。 圖12說明添加牛血清白蛋白至表面塗佈之QCM-D反應。 圖13為緩衝液沖洗之前及之後,表面塗佈上非特異性細胞(頂部圖像)及CTC(底部圖像)之像片。 圖14A說明各種表面塗佈之捕捉效率及非特異性血細胞結合。 圖14B為說明各種表面塗佈在緩衝液沖洗之前及之後的非特異性血細胞結合之像片。 圖15A至圖15C為在緩衝液沖洗純化之前及之後,表面塗佈上非特異性細胞及生物物質之像片。 圖16說明自表面塗佈移除HCT116及NIH-3T3細胞群體之不同剪切應力及沖洗時間。 圖17為由氣泡釋放的CTC之像片。 圖18說明所釋放之CTC在第1天、第10天及第14天之細胞培養物。 圖19圖解說明CTC過濾裝置。 圖20說明生物素標記之OC9801抗體、生物素標記之EpAb4-1抗體、生物素標記之EpCam抗體及IgG抗體之CTC結合特異性。
<110> 中央研究院 <120> 表面塗佈,使用表面塗佈捕捉、純化及釋放生物物質之方法與含有表面塗佈之新穎微流體晶片 <140> 101123391 <141> 2112-6-29 <150> US 61/502,844 <151> 2011-06-29 <150> US 61/606,220 <151> 2012-03-02 <160> 2 <170> FastSEQ for Windows Version 4.0 <210> 1 <211> 116 <212> PRT <213>人工序列 <220> <223> 合成之抗上皮細胞黏著分子膜(EpCAM)蛋白抗體EPAb4-1之重鏈V-H9結構域 <220> <221> 肽 <222> (1)...(25) <223> 構架區1 <220> <221> 肽 <222> (26)...(35) <223> 互補決定區3 <220> <221> 肽 <222> (36)...(50) <223> 構架區2 <220> <221> 肽 <222> (51)...(58) <223> 互補決定區2 <220> <221> 肽 <222> (59)...(98) <223> 構架區3 <220> <221> 肽 <222> (99)...(105) <223> 互補決定區3 <220> <221> 肽 <222> (106)...(116) <223> 構架區4 <400> 1 Gln Ile Gln Leu Val Gln Ser Gly Pro Glu Leu Lys Lys Pro Gly 1 5 10 15 Glu Thr Val Lys Ile Ser Cys Lys Ala Ser Gly Tyr Thr Phe Thr 20 25 30 Asn Tyr Gly Met Asn Trp Val Lys Gln Ala Pro Gly Lys Gly Leu 35 40 45 Lys Trp Met Gly Trp Ile Asn Thr Tyr Thr Gly Glu Pro Thr Tyr 50 55 60 Gly Asp Asp Phe Lys Gly Arg Phe Ala Phe Ser Leu Glu Thr Ser 65 70 75 Ala Ser Thr Ala Tyr Leu Gln Ile Asn Asn Leu Lys Asn Glu Asp 80 85 90 Thr Ala Thr Tyr Phe Cys Ala Arg Phe Gly Arg Ser Val Asp Phe 95 100 105 Trp Gly Gln Gly Thr Ser Val Thr Val Ser Ser 110 115 <210> 2 <211> 112 <212> PRT <213> 人工序列 <220> <223> 合成之抗上皮細胞黏著分子膜(EpCAM)蛋白抗體EPAb4-1之輕鏈V-kappa24/25結構域 <220> <221> 肽 <222> (1)...(23) <223> 構架區1 <220> <221>肽 <222> (24)...(39) <223> 互補決定區1 <220> <221> 肽 <222> (40)...(54) <223> 構架區2 <220> <221> 肽 <222> (55)...(61) <223> 互補決定區2 <220> <221> 肽 <222> (62)...(93) <223> 構架區3 <220> <221> 肽 <222> (94)...(102) <223> 互補決定區3 <220> <221> 肽 <222> (103)...(112) <223> 構架區4 <400> 2 Asp Ile Val Met Thr Gln Ala Ala Phe Ser Asn Pro Val Thr Leu 1 5 10 15 Gly Thr Ser Ala Ser Ile Ser Cys Arg Ser Ser Lys Ser Leu Leu 20 25 30 His Ser Asn Gly Ile Thr Tyr Leu Tyr Trp Tyr Leu Gln Lys Pro 35 40 45 Gly Gln Ser Pro Gln Leu Leu Ile Tyr His Met Ser Asn Leu Ala 50 55 60 Ser Gly Val Pro Asp Arg Phe Ser Ser Ser Gly Ser Gly Thr Asp 65 70 75 Phe Thr Leu Arg Ile Ser Arg Val Glu Ala Glu Asp Val Gly Ile 80 85 90 Tyr Tyr Cys Ala Gln Asn Leu Glu Asn Pro Arg Thr Phe Gly Gly 95 100 105 Gly Thr Lys Leu Glu Ile Lys 110
Claims (12)
- 一種方法,其包括:(a)使包含稀有細胞及非特異性細胞之生物樣本與微流裝置之非沾黏脂質層接觸,該微流裝置包含一抗體,該抗體選擇性地與稀有細胞結合,且其中該非沾黏脂質層係非共價連接該抗體;(b)使該等稀有細胞結合至該抗體;(c)使用液體洗滌該非沾黏脂質層,其中與該非沾黏脂質層接觸之非特異性細胞自該非沾黏脂質層釋放;以及(d)使包含氣泡之液體流過步驟(c)之微流裝置之通道,藉此自該微流裝置釋放該等稀有細胞。
- 如請求項1之方法,其中該經釋放的稀有細胞為活的。
- 如請求項2之方法,其進一步包括收集該等經釋放的稀有細胞。
- 如請求項1之方法,其中該抗體包含結合EpCAM之一重鏈及一輕鏈,其中:(a)該重鏈包含SEQ ID NO:1之CDR1、CDR2、及CDR3,及(b)該輕鏈包含SEQ ID NO:2之CDR1、CDR2、及CDR3。
- 如請求項1之方法,其中該非沾黏脂質層包含脂質多層或脂質體。
- 如請求項1之方法,其中該非沾黏脂質層包含脂質雙層。
- 如請求項1之方法,其中該非沾黏脂質層藉由表面連結分子連接於固體基板(solid substrate)。
- 如請求項1之方法,其中該非沾黏脂質層具有自幾奈米至幾百微米之厚度。
- 如請求項1之方法,其中該等稀有細胞為循環腫瘤細胞(CTCs)。
- 如請求項1之方法,其中該等非特異性細胞係藉由約2.5dyne/cm2至約10dyne/cm2之剪切應力選擇性地自該非沾黏脂質層被移除。
- 如請求項6之方法,其中該等氣泡內部之空氣與該脂質雙層之疏水性尾具有高親和力。
- 如請求項1之方法,其中該液體包含氣泡,其破壞該非沾黏脂質層且抬起該非沾黏脂質層脂質之頂層以及該等稀有細胞及經結合抗體。
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Country Status (5)
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Families Citing this family (47)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
EP2726870B1 (en) | 2011-06-29 | 2018-10-03 | Academia Sinica | The capture, purification and release of biological substance using a surface coating |
EP2755657B1 (en) | 2011-09-14 | 2017-11-29 | Neupharma, Inc. | Certain chemical entities, compositions, and methods |
US9977015B2 (en) * | 2012-05-15 | 2018-05-22 | Massachusetts Institute Of Technology | Systems and methods for detecting molecular interactions using magnetic beads |
US9494500B2 (en) | 2012-10-29 | 2016-11-15 | Academia Sinica | Collection and concentration system for biologic substance of interest and use thereof |
EP3126814B1 (en) * | 2014-04-01 | 2019-06-12 | Academia Sinica | Methods and systems for cancer diagnosis and prognosis |
WO2016019401A1 (en) * | 2014-07-30 | 2016-02-04 | Medvisionus Llc | Microfluidic device with smooth surface for enrichment of rare cells and biomarkers from a biological fluid |
CN105381824B (zh) * | 2014-08-26 | 2019-04-23 | 中央研究院 | 收集器架构布局设计 |
CN106170301A (zh) * | 2014-12-03 | 2016-11-30 | 士捷医疗设备(武汉)有限公司 | 一种模拟细胞体内环境的微流体器件及其应用 |
US20160178490A1 (en) | 2014-12-22 | 2016-06-23 | Saint-Gobain Performance Plastics Corporation | Capture system of cells and methods |
US10280390B2 (en) | 2014-12-22 | 2019-05-07 | Saint-Gobain Performance Plastics Corporation | System for culture of cells in a controlled environment |
US11175286B2 (en) | 2015-01-09 | 2021-11-16 | Spot Biosystems Ltd. | Immunolipoplex nanoparticle biochip containing molecular probes for capture and characterization of extracellular vesicles |
CN106148315B (zh) * | 2015-04-14 | 2019-02-01 | 中国科学院苏州纳米技术与纳米仿生研究所 | 一种基于壳聚糖纳米粒子的ctc捕获与纯化基底及其制备方法 |
SG11201708429WA (en) | 2015-04-22 | 2017-11-29 | Berkeley Lights Inc | Microfluidic cell culture |
EP3325966B1 (en) * | 2015-07-20 | 2021-01-20 | Sentilus Holdco, LLC | Chips, detectors, and methods of making and using the same |
US10799865B2 (en) | 2015-10-27 | 2020-10-13 | Berkeley Lights, Inc. | Microfluidic apparatus having an optimized electrowetting surface and related systems and methods |
TWI847100B (zh) | 2015-12-08 | 2024-07-01 | 美商布魯克細胞分析技術公司 | 在微流體器件中實施原位捕獲分析之套組 |
US10107726B2 (en) | 2016-03-16 | 2018-10-23 | Cellmax, Ltd. | Collection of suspended cells using a transferable membrane |
CN109346236A (zh) * | 2016-03-20 | 2019-02-15 | 李忠波 | 一种输电用防霉防菌抗腐蚀电缆 |
CN107287107A (zh) * | 2016-03-30 | 2017-10-24 | 无锡纳奥生物医药有限公司 | 一种循环肿瘤细胞分离设备、系统及方法 |
SG10202008265XA (en) | 2016-05-26 | 2020-09-29 | Berkeley Lights Inc | Covalently modified surfaces, kits, and methods of preparation and use |
CN107589254B (zh) * | 2016-07-08 | 2022-02-08 | 鉴识生物系统有限公司 | 一种免疫脂质体复合物纳米粒子生物芯片的制造方法及应用 |
US11085071B2 (en) | 2017-02-01 | 2021-08-10 | Spot Biosystems Ltd. | Highly stable and specific molecular beacons encapsulated in cationic lipoplex nanoparticles and application thereof |
US9738937B1 (en) | 2017-03-31 | 2017-08-22 | Cellmax, Ltd. | Identifying candidate cells using image analysis |
CN109097243A (zh) * | 2017-06-21 | 2018-12-28 | 曦医生技股份有限公司 | 生物微粒捕捉芯片组 |
CN107570482A (zh) * | 2017-07-06 | 2018-01-12 | 天津大学 | 界面的非特异性吸附物的去除装置及方法 |
TWI642685B (zh) * | 2017-07-26 | 2018-12-01 | 中原大學 | 新穎胜肽及其應用 |
WO2019025437A1 (en) | 2017-07-31 | 2019-02-07 | Universidad De Cantabria | CARTRIDGE, DEVICE AND METHOD FOR DETECTING, CAPTURING, IDENTIFYING AND COUNTING CIRCULATING TUMOR CELLS |
US11709156B2 (en) | 2017-09-18 | 2023-07-25 | Waters Technologies Corporation | Use of vapor deposition coated flow paths for improved analytical analysis |
US11709155B2 (en) | 2017-09-18 | 2023-07-25 | Waters Technologies Corporation | Use of vapor deposition coated flow paths for improved chromatography of metal interacting analytes |
CN107936085A (zh) * | 2017-11-21 | 2018-04-20 | 西南交通大学 | 利用紫外辐照在TiO2上固定蛋白并调控细胞亲和性的方法及TiO2‑蛋白产品 |
US11971414B2 (en) | 2017-12-22 | 2024-04-30 | Wisconsin Alumni Research Foundation | Nanoengineered surfaces for cancer biomarker capture |
WO2019161165A1 (en) | 2018-02-15 | 2019-08-22 | Ohio State Innovation Foundation | Microfluidic devices and methods for high throughput electroporation |
CN108795692B (zh) | 2018-06-26 | 2021-01-01 | 成都普瑞康生物科技有限公司 | 稀有细胞捕获系统及其应用 |
CN109187943B (zh) * | 2018-08-24 | 2021-07-30 | 四川新健康成生物股份有限公司 | 一种抗干扰试剂杯以及试剂杯内抗干扰涂层的制备方法 |
CN109136087A (zh) * | 2018-09-11 | 2019-01-04 | 京东方科技集团股份有限公司 | 分离芯片和分离方法 |
CN113195103A (zh) * | 2018-10-21 | 2021-07-30 | 堪萨斯大学 | 用于生成治疗递送平台的方法 |
CN109880148B (zh) * | 2019-01-22 | 2021-08-06 | 肇庆医学高等专科学校 | 一种表面印迹材料的制备及其在谷氨酸对映体拆分中的应用 |
CN110133267A (zh) * | 2019-06-18 | 2019-08-16 | 温州医科大学附属第二医院、温州医科大学附属育英儿童医院 | 一种用于吸附肿瘤细胞的载体,制备方法,试剂盒及应用 |
CN112300994B (zh) * | 2019-08-02 | 2024-01-12 | 中国科学院苏州纳米技术与纳米仿生研究所 | 用于捕获循环肿瘤细胞的纳米磁珠及其制备方法和应用 |
JP2023502629A (ja) * | 2019-11-26 | 2023-01-25 | ニクティア テクノロジーズ アーベー | 一般的な高容量タンパク質捕獲及び調節可能な電気化学的放出 |
US11918936B2 (en) | 2020-01-17 | 2024-03-05 | Waters Technologies Corporation | Performance and dynamic range for oligonucleotide bioanalysis through reduction of non specific binding |
US20240200141A1 (en) * | 2020-03-27 | 2024-06-20 | The Regents Of The University Of California | Covalent chemistry enables extracellular vesicle purification on nanosubstrates ? toward early detection of hepatocellular carcinoma |
CN111472200B (zh) * | 2020-05-19 | 2021-12-24 | 太原理工大学 | 一种纤维素试纸表面原位构建抗污凝胶涂层的方法 |
CN111632202A (zh) * | 2020-06-29 | 2020-09-08 | 南开大学 | 肿瘤细胞粘附材料及其制备方法和应用 |
CN112501122A (zh) * | 2020-12-09 | 2021-03-16 | 河南大学 | 一种肿瘤细胞的分离提取方法 |
CN112646635A (zh) * | 2020-12-22 | 2021-04-13 | 谢强 | 一种润滑油组合物及其制备方法 |
CN117043576A (zh) * | 2021-03-03 | 2023-11-10 | 华联生物科技股份有限公司 | 用于稀少细胞分离的生物电子系统及其应用 |
Family Cites Families (289)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US3784015A (en) | 1972-02-17 | 1974-01-08 | Bendix Corp | Filter |
US5147606A (en) | 1990-08-06 | 1992-09-15 | Miles Inc. | Self-metering fluid analysis device |
US5646001A (en) | 1991-03-25 | 1997-07-08 | Immunivest Corporation | Affinity-binding separation and release of one or more selected subset of biological entities from a mixed population thereof |
US5652148A (en) | 1993-04-20 | 1997-07-29 | Actimed Laboratories, Inc. | Method and apparatus for red blood cell separation |
US5427663A (en) | 1993-06-08 | 1995-06-27 | British Technology Group Usa Inc. | Microlithographic array for macromolecule and cell fractionation |
US5554686A (en) | 1993-08-20 | 1996-09-10 | Minnesota Mining And Manufacturing Company | Room temperature curable silane-terminated polyurethane dispersions |
US5707799A (en) | 1994-09-30 | 1998-01-13 | Abbott Laboratories | Devices and methods utilizing arrays of structures for analyte capture |
JP2000501184A (ja) | 1995-11-30 | 2000-02-02 | クロマビジョン メディカル システムズ,インコーポレイテッド | 生体標本の自動画像分析の方法および装置 |
US6718053B1 (en) | 1996-11-27 | 2004-04-06 | Chromavision Medical Systems, Inc. | Method and apparatus for automated image analysis of biological specimens |
US5885470A (en) | 1997-04-14 | 1999-03-23 | Caliper Technologies Corporation | Controlled fluid transport in microfabricated polymeric substrates |
US6890426B2 (en) | 1996-06-07 | 2005-05-10 | Immunivest Corporation | Magnetic separation apparatus and methods |
US6790366B2 (en) | 1996-06-07 | 2004-09-14 | Immunivest Corporation | Magnetic separation apparatus and methods |
US6074827A (en) | 1996-07-30 | 2000-06-13 | Aclara Biosciences, Inc. | Microfluidic method for nucleic acid purification and processing |
EP0929658A4 (en) | 1996-08-26 | 2005-11-02 | Univ Princeton | DEVICES FOR SORTING MICROSTRUCTURES THAT CAN BE REVERSIBLELY SEALED |
US6039897A (en) | 1996-08-28 | 2000-03-21 | University Of Washington | Multiple patterned structures on a single substrate fabricated by elastomeric micro-molding techniques |
DE69735601T2 (de) * | 1996-11-29 | 2007-10-18 | The Board Of Trustees Of The Leland Stanford Junior University, Stanford | Anordnungen von unabhängig voneinander ansteuerbaren, gestützten flüssigbilayer-membranen und ihre anwendungsverfahren |
US6583251B1 (en) * | 1997-09-08 | 2003-06-24 | Emory University | Modular cytomimetic biomaterials, transport studies, preparation and utilization thereof |
US5842787A (en) | 1997-10-09 | 1998-12-01 | Caliper Technologies Corporation | Microfluidic systems incorporating varied channel dimensions |
WO1999020649A1 (en) * | 1997-10-22 | 1999-04-29 | Merck Patent Gmbh | Spacer peptides and membranes containing same |
JP2002503814A (ja) | 1998-02-12 | 2002-02-05 | イムニベスト・コーポレイション | 循環ガン細胞の迅速かつ効率的な単離のための方法および試薬 |
US7282350B2 (en) | 1998-02-12 | 2007-10-16 | Immunivest Corporation | Labeled cell sets for use as functional controls in rare cell detection assays |
SE521415C2 (sv) | 1998-02-17 | 2003-10-28 | Hans Goeran Evald Martin | Metod för att framställa en gassensortillhörig detektor, samt en detektor framställd enligt metoden |
WO1999052574A1 (en) | 1998-04-10 | 1999-10-21 | Massachusetts Institute Of Technology | Biopolymers resistant coatings |
US6361749B1 (en) | 1998-08-18 | 2002-03-26 | Immunivest Corporation | Apparatus and methods for magnetic separation |
US20010036556A1 (en) | 1998-10-20 | 2001-11-01 | James S. Jen | Coatings for biomedical devices |
AU2876900A (en) | 1999-02-10 | 2000-08-29 | Cell Works Inc. | Class characterization of circulating cancer cells isolated from body fluids andmethods of use |
US6153113A (en) | 1999-02-22 | 2000-11-28 | Cobe Laboratories, Inc. | Method for using ligands in particle separation |
US6322683B1 (en) | 1999-04-14 | 2001-11-27 | Caliper Technologies Corp. | Alignment of multicomponent microfabricated structures |
AU5757100A (en) | 1999-06-21 | 2001-01-09 | General Hospital Corporation, The | Methods and devices for cell culturing and organ assist systems |
US6623982B1 (en) | 1999-07-12 | 2003-09-23 | Immunivest Corporation | Increased separation efficiency via controlled aggregation of magnetic nanoparticles |
US6790599B1 (en) | 1999-07-15 | 2004-09-14 | Microbionics, Inc. | Microfluidic devices and manufacture thereof |
US6271309B1 (en) | 1999-07-30 | 2001-08-07 | 3M Innovative Properties Company | Curable compositions comprising the hydrosilation product of olefin-containing polymers and organosiloxane hydrides, cured compositions made therefrom, and methods of making same |
ATE353920T1 (de) | 1999-12-27 | 2007-03-15 | Crucell Holland Bv | Menschlischer monoklonaler antikörper gegen ep- cam und dessen verwendung in krebstherapie |
JP3512775B2 (ja) | 2000-02-16 | 2004-03-31 | ウイスコンシン アラムニ リサーチ ファンデーション | 液晶アッセイ用の生化学的ブロッキング層 |
IL151873A0 (en) | 2000-03-24 | 2003-04-10 | Micromet Ag | Multifunctional polypeptides comprising a binding site to an epitope of the nkg2d receptor complex |
ES2264440T3 (es) | 2000-03-24 | 2007-01-01 | Micromet Ag | Amplificacion de mrna. |
US7769420B2 (en) | 2000-05-15 | 2010-08-03 | Silver James H | Sensors for detecting substances indicative of stroke, ischemia, or myocardial infarction |
WO2002004113A2 (en) | 2000-07-11 | 2002-01-17 | The Johns Hopkins University School Of Medicine | Methods of patterning protein and cell adhesivity |
CA2422162C (en) | 2000-09-09 | 2012-10-23 | The Research Foundation Of State University Of New York | Method and compositions for isolating metastatic cancer cells, and use in measuring metastatic potential of a cancer thereof |
NL1016779C2 (nl) | 2000-12-02 | 2002-06-04 | Cornelis Johannes Maria V Rijn | Matrijs, werkwijze voor het vervaardigen van precisieproducten met behulp van een matrijs, alsmede precisieproducten, in het bijzonder microzeven en membraanfilters, vervaardigd met een dergelijke matrijs. |
US6777836B2 (en) | 2000-12-20 | 2004-08-17 | General Electric Company | Heat transfer enhancement at generator stator core space blocks |
US6913697B2 (en) | 2001-02-14 | 2005-07-05 | Science & Technology Corporation @ Unm | Nanostructured separation and analysis devices for biological membranes |
AU2002251946A1 (en) | 2001-02-14 | 2002-08-28 | Science And Technology Corporation @ Unm | Nanostructured devices for separation and analysis |
US20060014013A1 (en) | 2001-03-10 | 2006-01-19 | Saavedra Steven S | Stabilized biocompatible supported lipid membrane |
WO2002081183A1 (en) | 2001-04-06 | 2002-10-17 | Fluidigm Corporation | Polymer surface modification |
PL205352B1 (pl) * | 2001-05-03 | 2010-04-30 | Merck Patent Gmbh | Rekombinowane przeciwciało specyficzne dla nowotworu |
US6844028B2 (en) | 2001-06-26 | 2005-01-18 | Accelr8 Technology Corporation | Functional surface coating |
US7501157B2 (en) | 2001-06-26 | 2009-03-10 | Accelr8 Technology Corporation | Hydroxyl functional surface coating |
US8815793B2 (en) | 2001-07-20 | 2014-08-26 | Northwestern University | Polymeric compositions and related methods of use |
WO2003008376A2 (en) | 2001-07-20 | 2003-01-30 | Northwestern University | Adhesive dopa-containing polymers and related methods of use |
US7858679B2 (en) | 2001-07-20 | 2010-12-28 | Northwestern University | Polymeric compositions and related methods of use |
US7863012B2 (en) | 2004-02-17 | 2011-01-04 | Veridex, Llc | Analysis of circulating tumor cells, fragments, and debris |
US20050230272A1 (en) | 2001-10-03 | 2005-10-20 | Lee Gil U | Porous biosensing device |
AU2002365079A1 (en) | 2001-10-03 | 2003-06-30 | Osman A. Basaran | Device and bioanalytical method utilizing asymmetric biofunction alized membrane |
US8986944B2 (en) | 2001-10-11 | 2015-03-24 | Aviva Biosciences Corporation | Methods and compositions for separating rare cells from fluid samples |
US8980568B2 (en) | 2001-10-11 | 2015-03-17 | Aviva Biosciences Corporation | Methods and compositions for detecting non-hematopoietic cells from a blood sample |
AU2002334989A1 (en) | 2001-10-11 | 2003-04-22 | Aviva Biosciences Corporation | Methods, compositions, and automated systems for separating rare cells from fluid samples |
EP1448807A4 (en) | 2001-10-30 | 2005-07-13 | Massachusetts Inst Technology | FLUORO CARBON ORGANOSILICIUM COPOLYMERS AND COATINGS MADE ACCORDING TO THE HFCVD PROCEDURE |
US20030163084A1 (en) | 2001-12-20 | 2003-08-28 | Klaus Tiemann | Creation and agitation of multi-component fluids in injection systems |
AU2003216175A1 (en) | 2002-02-04 | 2003-09-02 | Colorado School Of Mines | Laminar flow-based separations of colloidal and cellular particles |
GB0207350D0 (en) | 2002-03-28 | 2002-05-08 | Univ Sheffield | Surface |
US20050106570A1 (en) | 2002-04-03 | 2005-05-19 | Japan Science And Technology Agency | Biochip sensor surface carrying polyethylene glycolated nanoparticles |
US6955738B2 (en) | 2002-04-09 | 2005-10-18 | Gyros Ab | Microfluidic devices with new inner surfaces |
EP1572860B1 (en) | 2002-04-16 | 2018-12-05 | Princeton University | Gradient structures interfacing microfluidics and nanofluidics, methods for fabrication and uses thereof |
US7141369B2 (en) | 2002-04-25 | 2006-11-28 | Semibio Technology, Inc. | Measuring cellular metabolism of immobilized cells |
SE0201738D0 (sv) | 2002-06-07 | 2002-06-07 | Aamic Ab | Micro-fluid structures |
US7696320B2 (en) | 2004-08-24 | 2010-04-13 | Domantis Limited | Ligands that have binding specificity for VEGF and/or EGFR and methods of use therefor |
US8911831B2 (en) | 2002-07-19 | 2014-12-16 | Northwestern University | Surface independent, surface-modifying, multifunctional coatings and applications thereof |
WO2004011669A2 (en) | 2002-07-30 | 2004-02-05 | University Of Washington | Apparatus and methods for binding molecules and cells |
US20100233146A1 (en) | 2002-09-09 | 2010-09-16 | Reactive Surfaces, Ltd. | Coatings and Surface Treatments Having Active Enzymes and Peptides |
US20100210745A1 (en) | 2002-09-09 | 2010-08-19 | Reactive Surfaces, Ltd. | Molecular Healing of Polymeric Materials, Coatings, Plastics, Elastomers, Composites, Laminates, Adhesives, and Sealants by Active Enzymes |
US20040109853A1 (en) | 2002-09-09 | 2004-06-10 | Reactive Surfaces, Ltd. | Biological active coating components, coatings, and coated surfaces |
US20110240064A1 (en) | 2002-09-09 | 2011-10-06 | Reactive Surfaces, Ltd. | Polymeric Coatings Incorporating Bioactive Enzymes for Cleaning a Surface |
US20110250626A1 (en) | 2002-09-09 | 2011-10-13 | Reactive Surfaces, Ltd. | Visual Assays for Coatings Incorporating Bioactive Enzymes for Catalytic Functions |
JP2006501449A (ja) | 2002-09-27 | 2006-01-12 | ザ ジェネラル ホスピタル コーポレーション | 細胞分離のためのマイクロ流体デバイスおよびその使用 |
US8129330B2 (en) | 2002-09-30 | 2012-03-06 | Mountain View Pharmaceuticals, Inc. | Polymer conjugates with decreased antigenicity, methods of preparation and uses thereof |
DE10246069A1 (de) | 2002-10-02 | 2004-04-15 | Robert Bosch Gmbh | Vorrichtung zur Bestimmung wenigstens eines Parameters eines in einer Leitung strömenden Mediums |
AU2003299553A1 (en) | 2002-10-23 | 2004-05-13 | The Trustees Of Princeton University | Method for continuous particle separation using obstacle arrays asymmetrically aligned to fields |
EP1622691B1 (en) | 2003-03-14 | 2011-05-25 | The Trustees of Columbia University in the City of New York | Systems and methods of blood-based therapies having a microfluidic membraneless exchange device |
US20060076295A1 (en) | 2004-03-15 | 2006-04-13 | The Trustees Of Columbia University In The City Of New York | Systems and methods of blood-based therapies having a microfluidic membraneless exchange device |
US20040254419A1 (en) | 2003-04-08 | 2004-12-16 | Xingwu Wang | Therapeutic assembly |
US20070010702A1 (en) | 2003-04-08 | 2007-01-11 | Xingwu Wang | Medical device with low magnetic susceptibility |
US20050107870A1 (en) | 2003-04-08 | 2005-05-19 | Xingwu Wang | Medical device with multiple coating layers |
US20050079132A1 (en) | 2003-04-08 | 2005-04-14 | Xingwu Wang | Medical device with low magnetic susceptibility |
US20050025797A1 (en) | 2003-04-08 | 2005-02-03 | Xingwu Wang | Medical device with low magnetic susceptibility |
US7579077B2 (en) | 2003-05-05 | 2009-08-25 | Nanosys, Inc. | Nanofiber surfaces for use in enhanced surface area applications |
AU2004256392B2 (en) | 2003-04-28 | 2009-10-01 | Oned Material Llc | Super-hydrophobic surfaces, methods of their construction and uses therefor |
TWI427709B (zh) | 2003-05-05 | 2014-02-21 | Nanosys Inc | 用於增加表面面積之應用的奈米纖維表面 |
EP1636351A4 (en) | 2003-05-21 | 2007-04-04 | Gen Hospital Corp | MICROFABRICATED COMPOSITIONS AND METHOD FOR CONSTRUCTING TISSUES WITH MULTIPLE CELL TYPES |
GB0313569D0 (en) | 2003-06-12 | 2003-07-16 | Plasso Technology Ltd | Method |
US7544770B2 (en) * | 2003-08-29 | 2009-06-09 | Louisiana Tech Foundation, Inc. | Multilayer films, coatings, and microcapsules comprising polypeptides |
US7300801B2 (en) | 2003-09-12 | 2007-11-27 | 3M Innovative Properties Company | Welded sample preparation articles and methods |
US8101720B2 (en) | 2004-10-21 | 2012-01-24 | Xencor, Inc. | Immunoglobulin insertions, deletions and substitutions |
US8592219B2 (en) | 2005-01-17 | 2013-11-26 | Gyros Patent Ab | Protecting agent |
WO2007021762A2 (en) | 2005-08-09 | 2007-02-22 | The University Of North Carolina At Chapel Hill | Methods and materials for fabricating microfluidic devices |
US7117807B2 (en) | 2004-02-17 | 2006-10-10 | University Of Florida Research Foundation, Inc. | Dynamically modifiable polymer coatings and devices |
US9016221B2 (en) | 2004-02-17 | 2015-04-28 | University Of Florida Research Foundation, Inc. | Surface topographies for non-toxic bioadhesion control |
US20050181353A1 (en) | 2004-02-17 | 2005-08-18 | Rao Galla C. | Stabilization of cells and biological specimens for analysis |
US20060057180A1 (en) | 2004-02-20 | 2006-03-16 | Ashutosh Chilkoti | Tunable nonfouling surface of oligoethylene glycol |
SE0400662D0 (sv) | 2004-03-24 | 2004-03-24 | Aamic Ab | Assay device and method |
US20050215764A1 (en) | 2004-03-24 | 2005-09-29 | Tuszynski Jack A | Biological polymer with differently charged portions |
US7790458B2 (en) | 2004-05-14 | 2010-09-07 | Becton, Dickinson And Company | Material and methods for the growth of hematopoietic stem cells |
EP1744795A1 (en) | 2004-05-14 | 2007-01-24 | Becton, Dickinson and Company | Articles having bioactive surfaces and solvent-free methods of preparation thereof |
CA2567630A1 (en) | 2004-05-17 | 2005-11-24 | Mcmaster University | Biological molecule-reactive hydrophilic silicone surface |
JP2008501037A (ja) | 2004-06-01 | 2008-01-17 | マイクロチップス・インコーポレーテッド | 医療移植片への/医療移植片からの薬物または分析物の輸送の測定および輸送の増強のためのデバイスおよび方法 |
US20060251795A1 (en) | 2005-05-05 | 2006-11-09 | Boris Kobrin | Controlled vapor deposition of biocompatible coatings for medical devices |
US7695775B2 (en) | 2004-06-04 | 2010-04-13 | Applied Microstructures, Inc. | Controlled vapor deposition of biocompatible coatings over surface-treated substrates |
GB0420062D0 (en) | 2004-09-09 | 2004-10-13 | Akubio Ltd | Assay methods,materials and preparations |
US8663625B2 (en) | 2004-10-15 | 2014-03-04 | Cornell Research Foundation | Diffusively permeable monolithic biomaterial with embedded microfluidic channels |
WO2006050340A2 (en) | 2004-11-01 | 2006-05-11 | Polytechnic University | Methods and apparatus for modifying gel adhesion strength |
US20080274335A1 (en) | 2004-12-16 | 2008-11-06 | Regents Of The University Of Colorado | Photolytic Polymer Surface Modification |
US8069782B2 (en) | 2004-12-20 | 2011-12-06 | Nanoink, Inc. | Stamps with micrometer- and nanometer-scale features and methods of fabrication thereof |
SE0403139D0 (sv) | 2004-12-23 | 2004-12-23 | Nanoxis Ab | Device and use thereof |
US20090136982A1 (en) | 2005-01-18 | 2009-05-28 | Biocept, Inc. | Cell separation using microchannel having patterned posts |
US8158410B2 (en) | 2005-01-18 | 2012-04-17 | Biocept, Inc. | Recovery of rare cells using a microchannel apparatus with patterned posts |
CA2601918A1 (en) | 2005-01-20 | 2006-07-27 | The Regents Of The University Of California | Cellular microarrays for screening differentiation factors |
WO2006105313A2 (en) | 2005-03-29 | 2006-10-05 | Massachusetts Institute Of Technology | Compositions of and methods of using oversulfated glycosaminoglycans |
US20070026417A1 (en) | 2005-07-29 | 2007-02-01 | Martin Fuchs | Devices and methods for enrichment and alteration of circulating tumor cells and other particles |
US20070196820A1 (en) | 2005-04-05 | 2007-08-23 | Ravi Kapur | Devices and methods for enrichment and alteration of cells and other particles |
GB2472927B (en) | 2005-04-05 | 2011-05-04 | Gen Hospital Corp | Microfluidic Cell Capture on Micro-Corrugated Surface |
EP1874920A4 (en) | 2005-04-05 | 2009-11-04 | Cellpoint Diagnostics | DEVICES AND METHODS FOR ENRICHING AND MODIFYING CIRCULATING TUMOR CELLS AND OTHER PARTICLES |
US7485343B1 (en) | 2005-04-13 | 2009-02-03 | Sandia Corporation | Preparation of hydrophobic coatings |
US20060237390A1 (en) | 2005-04-14 | 2006-10-26 | King William P | Combined Microscale Mechanical Topography and Chemical Patterns on Polymer Substrates for Cell Culture |
US7846393B2 (en) | 2005-04-21 | 2010-12-07 | California Institute Of Technology | Membrane filter for capturing circulating tumor cells |
US7846743B2 (en) | 2005-04-21 | 2010-12-07 | California Institute Of Technology | Uses of parylene membrane filters |
CN100440572C (zh) * | 2005-05-31 | 2008-12-03 | 华南理工大学 | 一种有机小分子或高分子顶发射发光器件 |
US20070037173A1 (en) | 2005-08-12 | 2007-02-15 | Allard Jeffrey W | Circulating tumor cells (CTC's): early assessment of time to progression, survival and response to therapy in metastatic cancer patients |
SE0501418L (sv) | 2005-06-20 | 2006-09-26 | Aamic Ab | Metod och medel för att åstadkomma vätsketransport |
EP2004695A2 (en) | 2005-07-08 | 2008-12-24 | Xencor, Inc. | Optimized anti-ep-cam antibodies |
US7521195B1 (en) | 2005-07-21 | 2009-04-21 | Celera Corporation | Lung disease targets and uses thereof |
US8088161B2 (en) | 2005-07-28 | 2012-01-03 | Visioncare Ophthalmic Technologies Inc. | Compressed haptics |
US8921102B2 (en) | 2005-07-29 | 2014-12-30 | Gpb Scientific, Llc | Devices and methods for enrichment and alteration of circulating tumor cells and other particles |
US20070026416A1 (en) | 2005-07-29 | 2007-02-01 | Martin Fuchs | Devices and methods for enrichment and alteration of circulating tumor cells and other particles |
US7431969B2 (en) | 2005-08-05 | 2008-10-07 | Massachusetts Institute Of Technology | Chemical vapor deposition of hydrogel films |
US7993821B2 (en) | 2005-08-11 | 2011-08-09 | University Of Washington | Methods and apparatus for the isolation and enrichment of circulating tumor cells |
US7901950B2 (en) | 2005-08-12 | 2011-03-08 | Veridex, Llc | Method for assessing disease states by profile analysis of isolated circulating endothelial cells |
EP1931718B1 (en) | 2005-08-24 | 2012-12-26 | ETH Zurich | Catechol functionalized polymers and method for preparing them |
US20110097277A1 (en) | 2005-08-25 | 2011-04-28 | University Of Washington | Particles coated with zwitterionic polymers |
CN101341243A (zh) | 2005-08-25 | 2009-01-07 | 索利克斯生物燃料公司 | 由藻类生产生物柴油的方法、设备和系统 |
WO2007024393A2 (en) | 2005-08-25 | 2007-03-01 | University Of Washington | Super-low fouling sulfobetaine and carboxybetaine materials and related methods |
US20070059716A1 (en) | 2005-09-15 | 2007-03-15 | Ulysses Balis | Methods for detecting fetal abnormality |
WO2007035527A2 (en) | 2005-09-15 | 2007-03-29 | Duke University | Non-fouling polymeric surface modification and signal amplification method for biomolecular detection |
WO2007035842A2 (en) | 2005-09-21 | 2007-03-29 | Ccc Diagnostics, Llc | Comprehensive diagnostic testing procedures for personalized anticancer chemotherapy (pac) |
US7846445B2 (en) | 2005-09-27 | 2010-12-07 | Amunix Operating, Inc. | Methods for production of unstructured recombinant polymers and uses thereof |
US7855279B2 (en) | 2005-09-27 | 2010-12-21 | Amunix Operating, Inc. | Unstructured recombinant polymers and uses thereof |
WO2007044616A2 (en) | 2005-10-06 | 2007-04-19 | Xencor, Inc. | Optimized anti-cd30 antibodies |
ES2341886T3 (es) * | 2005-10-28 | 2010-06-29 | Max-Planck-Gesellschaft Zur Forderung Der Wissenschaften E.V. | Transcripcion y traduccion in vitro libre de celulas de proteinas de membrana en capas lipidicas planas unidas. |
JP5129149B2 (ja) | 2005-10-31 | 2013-01-23 | ザ リージェンツ オブ ザ ユニバーシティ オブ ミシガン | 癌を処置および診断するための組成物および方法 |
WO2007056561A2 (en) | 2005-11-09 | 2007-05-18 | Liquidia Technologies, Inc. | Medical device, materials, and methods |
KR100763907B1 (ko) * | 2005-12-26 | 2007-10-05 | 삼성전자주식회사 | 미세유동 장치의 제조방법 및 그에 의하여 제조되는미세유동 장치 |
WO2007079229A2 (en) | 2005-12-29 | 2007-07-12 | Cellpoint Diagnostics, Inc. | Devices and methods for enrichment and alteration of circulating tumor cells and other particles |
US20090317836A1 (en) | 2006-01-30 | 2009-12-24 | The Scripps Research Institute | Methods for Detection of Circulating Tumor Cells and Methods of Diagnosis of Cancer in Mammalian Subject |
WO2007089762A2 (en) | 2006-01-31 | 2007-08-09 | Biomed Solutions, Llc | Devices for selective recruitment, isolation, activation, and/or elimination of various cell populations |
WO2007090779A1 (en) | 2006-02-10 | 2007-08-16 | Universite De Liege | Electrografting method for forming and regulating a strong adherent nanostructured polymer coating |
GB2448858A (en) | 2006-02-15 | 2008-11-05 | Aida Eng Ltd | Microchannel chip and method for manufacturing such chip |
DE102006009004A1 (de) | 2006-02-23 | 2007-09-06 | Sustech Gmbh & Co. Kg | Multifunktionelle sternförmige Präpolymere, deren Herstellung und Verwendung |
US20080213853A1 (en) | 2006-02-27 | 2008-09-04 | Antonio Garcia | Magnetofluidics |
CA2646293C (en) | 2006-03-15 | 2015-08-25 | The General Hospital Corporation | Devices and methods for detecting cells and other analytes |
WO2007121465A2 (en) | 2006-04-18 | 2007-10-25 | Wellstat Biologics Corporation | Detection of proteins from circulating neoplastic cells |
US8212083B2 (en) | 2006-04-27 | 2012-07-03 | Intezyne Technologies, Inc. | Heterobifunctional poly(ethylene glycol) containing acid-labile amino protecting groups and uses thereof |
EP2010477A4 (en) | 2006-04-27 | 2012-05-30 | Intezyne Technologies Inc | POLY (ETHYLENE GLYCOL) CONTAINING CHEMICALLY DISPARATE ENDOGROUP |
CA2652027C (en) | 2006-05-12 | 2017-06-06 | Immunivest Corporation | A laser illumination system in fluorescent microscopy |
AU2007253702A1 (en) | 2006-05-22 | 2007-11-29 | The Trustees Of Columbia University In The City Of New York | Systems and methods of microfluidic membraneless exchange using filtration of extraction fluid outlet streams |
EP2040843B1 (en) | 2006-06-01 | 2020-02-26 | The Trustees of Princeton University | Apparatus for continuous particle separation |
CA2675596A1 (en) | 2006-06-06 | 2007-12-13 | Societe De Commercialisation Des Produits De La Recherche Appliquee - So Cpra-Sciences Et Genie S.E.C. | Assay supports comprising a peg support, said support attached from a peg solution in cloud point (theta solvent) conditions |
US20080090239A1 (en) | 2006-06-14 | 2008-04-17 | Daniel Shoemaker | Rare cell analysis using sample splitting and dna tags |
US8535791B2 (en) | 2006-06-30 | 2013-09-17 | The University Of Akron | Aligned carbon nanotube-polymer materials, systems and methods |
CN101583722A (zh) | 2006-07-14 | 2009-11-18 | 阿维瓦生物科学股份有限公司 | 从生物学样品检测稀有细胞的方法和组合物 |
EP2069412B1 (en) | 2006-08-07 | 2019-01-02 | University of Washington | Mixed charge copolymers and hydrogels |
AU2007286203B2 (en) | 2006-08-08 | 2013-05-02 | Board Of Regents, The University Of Texas System | Multistage delivery of active agents |
US8114431B2 (en) | 2006-09-19 | 2012-02-14 | Georgia Tech Research Corporation | Biomolecular coating for implants |
GB0618460D0 (en) | 2006-09-20 | 2006-11-01 | Univ Belfast | Process for preparing surfaces with tailored wettability |
EP2078062B1 (en) | 2006-10-19 | 2018-12-05 | Northwestern University | Surface-independent, surface-modifying, multifunctional coatings and applications thereof |
WO2008052358A1 (en) | 2006-11-03 | 2008-05-08 | The Governors Of The University Of Alberta | Microfluidic device having an array of spots |
US20080114096A1 (en) | 2006-11-09 | 2008-05-15 | Hydromer, Inc. | Lubricious biopolymeric network compositions and methods of making same |
US20080113350A1 (en) | 2006-11-09 | 2008-05-15 | Terstappen Leon W M M | Blood test to monitor the genetic changes of progressive cancer using immunomagnetic enrichment and fluorescence in situ hybridization (FISH) |
GB0623160D0 (en) | 2006-11-20 | 2006-12-27 | Smith & Nephew | Biomolecules |
EP2097119A4 (en) | 2006-11-21 | 2012-10-17 | Abbott Lab | USE OF A TERPOLYMER OF TETRAFLUOROETHYLENE, HEXAFLUORPROPYLENE AND VINYLIDENE FLUORIDE IN MEDICAMENTAL COATINGS |
FR2909013B1 (fr) | 2006-11-28 | 2011-02-25 | Commissariat Energie Atomique | Procede de revetement en film mince. |
JP2010530955A (ja) | 2006-12-29 | 2010-09-16 | ユニヴァーシティ オブ ワシントン | 二重機能性非汚染表面および材料 |
WO2008089282A2 (en) | 2007-01-16 | 2008-07-24 | Silver James H | Sensors for detecting subtances indicative of stroke, ischemia, infection or inflammation |
US8057418B2 (en) | 2007-03-01 | 2011-11-15 | Nanospectra Biosciences, Inc. | Devices and methods for extracorporeal ablation of circulating cells |
US7981688B2 (en) | 2007-03-08 | 2011-07-19 | University Of Washington | Stimuli-responsive magnetic nanoparticles and related methods |
WO2008121375A2 (en) | 2007-03-29 | 2008-10-09 | Pacific Biosciences Of California, Inc. | Modified surfaces for immobilization of active molecules |
ES2367675T3 (es) | 2007-04-04 | 2011-11-07 | Sigma-Tau Industrie Farmaceutiche Riunite S.P.A. | Anticuerpo anti-epcam y usos del mismo. |
EP2482055A3 (en) | 2007-04-16 | 2013-10-30 | The General Hospital Corporation d/b/a Massachusetts General Hospital | Systems and methods for particle focusing in microchannels |
US20100233693A1 (en) | 2007-04-16 | 2010-09-16 | On-O-ity, Inc | Methods for diagnosing, prognosing, or theranosing a condition using rare cells |
US20100112026A1 (en) * | 2007-04-18 | 2010-05-06 | Massachusetts Institute To Technology | Surfaces, methods and devices employing cell rolling |
US8945909B2 (en) | 2007-04-25 | 2015-02-03 | The Regents Of The University Of Michigan | Tunable elastomeric nanochannels for nanofluidic manipulation |
WO2009009185A2 (en) | 2007-05-09 | 2009-01-15 | Massachusetts Institute Of Technology | Tunable surfaces |
JP5067588B2 (ja) | 2007-05-18 | 2012-11-07 | 富士レビオ株式会社 | 表面−固定化生物学的分子の活性増大のための化学的表面ナノパターン |
US8063187B2 (en) | 2007-05-30 | 2011-11-22 | Xencor, Inc. | Methods and compositions for inhibiting CD32B expressing cells |
US20080312356A1 (en) | 2007-06-13 | 2008-12-18 | Applied Mcrostructures, Inc. | Vapor-deposited biocompatible coatings which adhere to various plastics and metal |
US8841135B2 (en) | 2007-06-20 | 2014-09-23 | University Of Washington | Biochip for high-throughput screening of circulating tumor cells |
WO2009009408A2 (en) | 2007-07-06 | 2009-01-15 | Applied Biosystems Inc. | Devices and methods for the detection of analytes |
EP2020261A1 (en) | 2007-07-20 | 2009-02-04 | Nederlandse Organisatie voor toegepast- natuurwetenschappelijk onderzoek TNO | Multi component particle generating system |
DE102007039665A1 (de) | 2007-08-22 | 2009-02-26 | Sustech Gmbh & Co. Kg | Silylfunktionelle lineare Präpolymere, deren Herstellung und Verwendung |
EP2037265A1 (en) | 2007-09-17 | 2009-03-18 | Adnagen AG | Solid phase cell isolation and/or enrichment method |
US7736891B2 (en) | 2007-09-11 | 2010-06-15 | University Of Washington | Microfluidic assay system with dispersion monitoring |
US11307201B2 (en) | 2007-09-17 | 2022-04-19 | Red Ivory Llc | Self-actuating signal producing detection devices and methods |
US9150788B2 (en) | 2007-09-18 | 2015-10-06 | Syracuse University | Non-amphiphile-based water-in-water emulsion and uses thereof |
WO2009079053A2 (en) | 2007-09-19 | 2009-06-25 | Massachusetts Institute Of Technology | High affinity metal-oxide binding peptides with reversible binding |
WO2009043057A2 (en) | 2007-09-27 | 2009-04-02 | Massachusetts Institute Of Technology | Cell rolling separation |
US20090098017A1 (en) | 2007-10-16 | 2009-04-16 | Board Of Regents, The University Of Texas System | Nanoporous membrane exchanger |
WO2009051734A1 (en) | 2007-10-17 | 2009-04-23 | The General Hospital Corporation | Microchip-based devices for capturing circulating tumor cells and methods of their use |
WO2009061787A1 (en) | 2007-11-05 | 2009-05-14 | Nanocopoeia, Inc. | Coated devices and method of making coated devices that reduce smooth muscle cell proliferation and platelet activity |
AU2008326438B2 (en) | 2007-11-19 | 2014-09-04 | University Of Washington | Cationic betaine precursors to zwitterionic betaines having controlled biological properties |
US8658192B2 (en) | 2007-11-19 | 2014-02-25 | University Of Washington | Integrated antimicrobial and low fouling materials |
US20090181441A1 (en) | 2007-11-27 | 2009-07-16 | Board Of Trustees Of Michigan State University | Porous silicon-polymer composites for biosensor applications |
CN202011883U (zh) | 2007-12-12 | 2011-10-19 | 里兰斯坦福初级大学理事会 | 用于捕获和分离目标细胞的装置 |
WO2009077760A1 (en) | 2007-12-17 | 2009-06-25 | Lux Innovate Limited | Compositions and methods for maintenance of fluid conducting and containment systems |
US20110008404A1 (en) | 2007-12-19 | 2011-01-13 | Georgia Tech Research Corporation | Modification Of Biomaterials With Microgel Films |
EP2980217A1 (en) * | 2007-12-31 | 2016-02-03 | XOMA Technology Ltd. | Methods and materials for targeted mutagenesis |
GB0801600D0 (en) | 2008-01-29 | 2008-03-05 | Univ Cardiff | Microtrench |
CA2714594A1 (en) | 2008-02-04 | 2009-08-13 | Edward F. Leonard | Fluid separation devices, systems and methods |
US20100323918A1 (en) | 2008-02-10 | 2010-12-23 | Microdysis, Inc | Polymer surface functionalization and related applications |
CA2639954C (en) | 2008-02-11 | 2017-08-15 | Aaron R. Wheeler | Droplet-based cell culture and cell assays using digital microfluidics |
US8008032B2 (en) | 2008-02-25 | 2011-08-30 | Cellective Dx Corporation | Tagged ligands for enrichment of rare analytes from a mixed sample |
WO2009111159A2 (en) | 2008-03-03 | 2009-09-11 | New York University | Biocompatible materials containing stable complexes of tsg-6 and hyaluronan and method of using same |
ES2968853T3 (es) | 2008-03-24 | 2024-05-14 | Univ Louisiana State | Aislamiento microfluídico de células tumorales u otras células raras a partir de sangre total u otros líquidos |
US8796184B2 (en) | 2008-03-28 | 2014-08-05 | Sentilus, Inc. | Detection assay devices and methods of making and using the same |
WO2009120151A1 (en) | 2008-03-28 | 2009-10-01 | Nanyang Technological University | Membrane made of a nanostructured material |
US8128983B2 (en) | 2008-04-11 | 2012-03-06 | Abbott Cardiovascular Systems Inc. | Coating comprising poly(ethylene glycol)-poly(lactide-glycolide-caprolactone) interpenetrating network |
US8567612B2 (en) | 2008-04-15 | 2013-10-29 | Nanoh2O, Inc. | Hybrid TFC RO membranes with nitrogen additives |
US8177978B2 (en) | 2008-04-15 | 2012-05-15 | Nanoh20, Inc. | Reverse osmosis membranes |
SE533515C2 (sv) | 2008-04-16 | 2010-10-12 | Aamic Ab | Analysförfarande för samtidig analys av flera analyter |
US20090285873A1 (en) | 2008-04-18 | 2009-11-19 | Abbott Cardiovascular Systems Inc. | Implantable medical devices and coatings therefor comprising block copolymers of poly(ethylene glycol) and a poly(lactide-glycolide) |
US8916188B2 (en) | 2008-04-18 | 2014-12-23 | Abbott Cardiovascular Systems Inc. | Block copolymer comprising at least one polyester block and a poly (ethylene glycol) block |
US20090264317A1 (en) | 2008-04-18 | 2009-10-22 | University Of Massachusetts | Functionalized nanostructure, methods of manufacture thereof and articles comprising the same |
JP5042110B2 (ja) | 2008-04-22 | 2012-10-03 | サルナス、ペトロニス | ナノ細孔の製造 |
GB0810039D0 (en) | 2008-06-03 | 2008-07-09 | Univ Belfast | Shape-formed product with tailored wettability |
US8833430B2 (en) | 2008-06-26 | 2014-09-16 | President And Fellows Of Harvard College | Versatile high aspect ratio actuatable nanostructured materials through replication |
WO2010123608A2 (en) * | 2009-01-29 | 2010-10-28 | The Regents Of The University Of California | A spatial biomarker of disease and detection of spatial organization of cellular recptors |
US8579117B2 (en) | 2008-07-24 | 2013-11-12 | The Trustees Of Princeton University | Bump array device having asymmetric gaps for segregation of particles |
CN101903174B (zh) | 2008-08-11 | 2014-04-02 | 北京大学 | 超疏水聚二甲基硅氧烷及其制备方法 |
US20100055733A1 (en) | 2008-09-04 | 2010-03-04 | Lutolf Matthias P | Manufacture and uses of reactive microcontact printing of biomolecules on soft hydrogels |
WO2010028160A1 (en) | 2008-09-05 | 2010-03-11 | The Scripps Research Institute | Methods for the detection of circulating tumor cells |
US8367090B2 (en) | 2008-09-05 | 2013-02-05 | Abbott Cardiovascular Systems Inc. | Coating on a balloon comprising a polymer and a drug |
US20100096327A1 (en) | 2008-09-19 | 2010-04-22 | Gin Douglas L | Polymer coatings that resist adsorption of proteins |
WO2010034484A1 (en) | 2008-09-23 | 2010-04-01 | Gilupi Gmbh | Diagnostic analyte collection device based on flexible polymers with biological surface modification and microfluidic functionality |
EP2344625A4 (en) | 2008-10-02 | 2012-02-08 | Univ California | HIGH RESOLUTION MICROSTRUCTURING METHODS AND COMPOSITIONS FOR CELL CULTURE |
US20110212440A1 (en) | 2008-10-10 | 2011-09-01 | Cnrs-Dae | Cell sorting device |
US8535644B2 (en) | 2008-10-10 | 2013-09-17 | Massachusetts Institute Of Technology | Tunable hydrogel microparticles |
WO2010053993A1 (en) | 2008-11-04 | 2010-05-14 | The Trustees Of Columbia University In The City Of New York | Heterobifunctional polymers and methods for layer-by-layer construction of multilayer films |
US8292492B2 (en) | 2008-11-11 | 2012-10-23 | Sandia Corporation | Airfoil-shaped micro-mixers for reducing fouling on membrane surfaces |
EP2352796A1 (en) | 2008-12-05 | 2011-08-10 | Semprus Biociences Corporation | Layered non-fouling, antimicrobial, antithrombogenic coatings |
KR20110106866A (ko) | 2008-12-05 | 2011-09-29 | 셈프러스 바이오사이언시스 코퍼레이션 | 비-파울링, 항-미생물성, 항-혈전형성성 그라프트-프롬 조성물 |
CA2745926A1 (en) | 2008-12-08 | 2010-07-08 | Phaserx, Inc. | Omega-functionalized polymers, junction-functionalized block copolymers, polymer bioconjugates, and radical chain extension polymerization |
JP2012519558A (ja) | 2009-03-10 | 2012-08-30 | モナッシュ ユニヴァーシティ | マイクロ流体素子を使用する血小板凝集 |
US9140697B2 (en) | 2009-03-18 | 2015-09-22 | The Regents Of The University Of California | Device for capturing circulating cells |
EP2412020B1 (en) | 2009-03-24 | 2020-09-30 | University Of Chicago | Slip chip device and methods |
US8343440B2 (en) | 2009-03-27 | 2013-01-01 | Seiko Epson Corporation | Cell separating apparatus and cell separating method |
US8569463B2 (en) | 2009-04-23 | 2013-10-29 | Syracuse University | Method of covalently modifying proteins with organic molecules to prevent aggregation |
EP2421628A4 (en) | 2009-04-23 | 2012-10-10 | Logos Energy Inc | LATERAL DISPLACEMENT ARRANGEMENT FOR MICROFILTRATION |
US20120077246A1 (en) | 2009-04-24 | 2012-03-29 | The Board Of Trustees Of The University Of Illinoi | Methods and Devices for Capturing Circulating Tumor Cells |
US20120270209A1 (en) | 2009-05-15 | 2012-10-25 | Massachusetts Institute Of Technology | Systems, devices, and methods for specific capture and release of biological sample components |
WO2010147918A2 (en) | 2009-06-15 | 2010-12-23 | 3M Innovative Properties Company | Detection of acid-producing bacteria |
JP2013504064A (ja) | 2009-09-03 | 2013-02-04 | ザ スクリプス リサーチ インスティチュート | 循環腫瘍細胞を分類する方法 |
US8481336B2 (en) | 2009-09-09 | 2013-07-09 | The Board Of Trustees Of The Leland Stanford Junior University | Magnetic separation device for cell sorting and analysis |
CA2773907A1 (en) | 2009-09-21 | 2011-03-24 | Mount Sinai Hospital | Methods and compositions for the diagnosis and treatment of thyroid cancer |
CN102803969A (zh) | 2009-09-25 | 2012-11-28 | 傲锐东源生物科技有限公司 | 蛋白质阵列及其用途 |
CN101701039B (zh) * | 2009-11-06 | 2011-10-26 | 中国人民解放军第四军医大学 | Fmu-epcam-2a9单克隆抗体的轻链和重链可变区 |
US20110165161A1 (en) | 2009-12-23 | 2011-07-07 | Shih-Yao Lin | Anti-epcam antibodies that induce apoptosis of cancer cells and methods using same |
WO2011112969A1 (en) | 2010-03-11 | 2011-09-15 | Intezyne Technologies, Inc. | Poly(ethylene glycol) derivatives for metal-free click chemistry |
AU2011250588C1 (en) | 2010-05-04 | 2016-09-01 | Ranju Ralhan | Method for the diagnosis of epithelial cancers by the detection of EpICD polypeptide |
TWI539158B (zh) | 2010-06-08 | 2016-06-21 | 維里德克斯有限責任公司 | 使用血液中之循環黑色素瘤細胞預測黑色素瘤病患之臨床結果的方法。 |
CA2799786A1 (en) | 2010-06-09 | 2011-12-15 | Semprus Biosciences Corp. | Non-fouling, anti-microbial, anti-thrombogenic graft compositions |
JP6083071B2 (ja) | 2010-06-09 | 2017-02-22 | アロー インターナショナル インコーポレイテッド | 非汚染性、抗菌性、抗血栓性グラフトフロム組成物 |
US8574660B2 (en) | 2010-06-09 | 2013-11-05 | Semprus Biosciences Corporation | Articles having non-fouling surfaces and processes for preparing the same without altering bulk physical properties |
US9248467B2 (en) | 2010-07-13 | 2016-02-02 | Rigoberto Advincula | Types of electrodeposited polymer coatings with reversible wettability and electro-optical properties |
WO2012016136A2 (en) | 2010-07-30 | 2012-02-02 | The General Hospital Corporation | Microscale and nanoscale structures for manipulating particles |
MX2013001750A (es) | 2010-08-15 | 2013-06-05 | Gpb Scientific Llc | Separacion celular micro-fluidica en el ensayo de sangre. |
WO2012031201A2 (en) | 2010-09-03 | 2012-03-08 | Massachusetts Institute Of Technology | Fabrication of anti-fouling surfaces comprising a micro- or nano-patterned coating |
JP5920895B2 (ja) | 2010-09-14 | 2016-05-25 | ザ リージェンツ オブ ザ ユニバーシティ オブ カリフォルニア | マイクロ流体捕獲渦を使用して不均一溶液から細胞を単離する方法及びデバイス |
CN102011193B (zh) | 2010-09-21 | 2013-03-13 | 南京航空航天大学 | 蛋白质修饰的GaN纳米线阵列及其制法和用途 |
US20140154703A1 (en) | 2011-01-06 | 2014-06-05 | Alison Skelley | Circulating tumor cell capture on a microfluidic chip incorporating both affinity and size |
WO2012103025A2 (en) | 2011-01-24 | 2012-08-02 | Epic Sciences, Inc. | Methods for obtaining single cells and applications of single cell omics |
US9212258B2 (en) | 2011-02-23 | 2015-12-15 | The Board Of Trustees Of The University Of Illinois | Amphiphilic dendron-coils, micelles thereof and uses |
USD650091S1 (en) | 2011-04-19 | 2011-12-06 | Zach Odeh | Microfluidic device |
KR101279918B1 (ko) | 2011-05-27 | 2013-07-01 | 한국과학기술연구원 | 종양세포 검출장치 및 종양세포 검출방법 |
EP2726870B1 (en) | 2011-06-29 | 2018-10-03 | Academia Sinica | The capture, purification and release of biological substance using a surface coating |
US20130157347A1 (en) | 2011-07-07 | 2013-06-20 | Veridex, Llc | Method of Analyzing Cardiovascular Disorders and Uses Thereof |
JP6133297B2 (ja) | 2011-09-06 | 2017-05-24 | ベクトン・ディキンソン・アンド・カンパニーBecton, Dickinson And Company | 試料中の希少標的細胞のサイトメトリー検出のための方法及び組成物 |
US20140296095A1 (en) | 2011-09-23 | 2014-10-02 | The Trustees Of Columbia University In The City Of New York | Spatially Selective Release of Aptamer-Captured Cells by Temperature Mediation |
JP6163502B2 (ja) | 2012-03-02 | 2017-07-12 | アカデミア シニカAcademia Sinica | 抗上皮細胞接着分子(EpCAM)抗体及びその使用方法 |
US9494500B2 (en) | 2012-10-29 | 2016-11-15 | Academia Sinica | Collection and concentration system for biologic substance of interest and use thereof |
EP3126814B1 (en) | 2014-04-01 | 2019-06-12 | Academia Sinica | Methods and systems for cancer diagnosis and prognosis |
CN105381824B (zh) | 2014-08-26 | 2019-04-23 | 中央研究院 | 收集器架构布局设计 |
US10107726B2 (en) | 2016-03-16 | 2018-10-23 | Cellmax, Ltd. | Collection of suspended cells using a transferable membrane |
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- 2012-06-28 CN CN201710536863.4A patent/CN107315086B/zh active Active
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EP2726870A2 (en) | 2014-05-07 |
US20140255976A1 (en) | 2014-09-11 |
TW201636593A (zh) | 2016-10-16 |
CN110763842A (zh) | 2020-02-07 |
CN103998932A (zh) | 2014-08-20 |
WO2013003624A2 (en) | 2013-01-03 |
US9541480B2 (en) | 2017-01-10 |
TWI550273B (zh) | 2016-09-21 |
EP2726870B1 (en) | 2018-10-03 |
CN107315086A (zh) | 2017-11-03 |
US11674958B2 (en) | 2023-06-13 |
US20210088514A1 (en) | 2021-03-25 |
CN107315086B (zh) | 2019-09-10 |
TW201323875A (zh) | 2013-06-16 |
CN103998932B (zh) | 2017-06-06 |
WO2013003624A3 (en) | 2013-03-14 |
EP2726870A4 (en) | 2015-03-11 |
US20170199184A1 (en) | 2017-07-13 |
US20230366879A1 (en) | 2023-11-16 |
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