1275797 A7 B7 五、發明説明（J ) 技術領域 本發明關於可被用於製造生物晶片之塑膠載片。詳言 之’本發明關於具有特殊構造之經模製塑膠載片，其可被 用於製備表面經化學處理之塑膠載片，該經處理之表面上 可被固定微陣列之生物樣品並形成斑點，因而製造生物晶 片。 發明背景 玻璃載片已被廣泛地使用，以作爲製造D N A晶片和 蛋白質晶片時所需之基材。然而，迄今使用玻璃載片於作 爲生物晶片之基材時，已顯現出若干缺點。首先，玻璃載 片本身易碎，且於操作上需小心處理。玻璃載片之表面不 易經化學處理以期能成功地固定生物性小分子化合物（諸 如，植物或草藥之代謝物）至該經處理之表面上以達到直 接結合之目的。玻璃載片亦產生不欲之高背景訊號，其在 對隨後經標記物處理之生物晶片作分析時會產生干擾。因 此，此技藝需要開發出新材料或發展出能夠易於修飾或改 質之材料，以取代玻璃材料作爲生物晶片之基材。 近來，已對開發塑膠載片爲基材之生物晶片作出某些 努力，作爲此晶片基材之塑膠載片不僅比玻璃載片之取得 成本較低，且亦可藉由慣用之注射模塑法輕易地模塑成所 欲之形狀。 w 0 0 0 / 5 5 6 2 7揭示一種用於偵測標的分析 物之生物晶片，其包含固定於非螢光之丙烯酸性基材上之 本紙張尺度適用中國國家標準（CNS ) A4規格（210 X 297公釐） K - n^— m —ϋΜ mu ·Γϋ m· mi -......- (請先閱讀背面之注意事項再填寫本頁) 訂 ΦΓ 經濟部智慧財產局員工消費合作社印製 -4- 1275797 A7 _______ B7 ___ 五、發明説明（2 ) 一列生物性結合配體。 W 0 0 0 / 3 6 1 4 5揭示一種製造生物晶片之方 (請先閱讀背面之注意事項再填寫本頁) 法，其包含接枝生物性探針至導電性聚合物上。 EP 1 026 259揭示一種DNA晶片，其 包含固體載體及於親水性聚合物之存在下固定於該固體載 體上之寡核苷酸或聚核苷酸。 雖然塑膠材質已被使用以製造D N A晶片，然而迄今 所使用之塑膠載片皆爲平坦的，亦即在上方表面上並無任 何凹槽或突出物，且其會產生不欲之高背景訊號，其在對 生物晶片作分析時會產生干擾。迄今並未成功地使用表面 經處理之塑膠載片以固定（適宜地是以微陣列之形式）蛋 白質、肽或小分子化合物至其經處理之表面上，特別是在 該蛋白質、肽或小分子化合物未經修飾之情況下。 此外，先前技藝並未教示或建議聚苯乙烯材質之塑膠 載片的表面可藉由利用簡單之試劑以一步驟之方式加以處 理’使得寡核苷酸或聚核苷酸能夠固定（適宜地是以微陣 列之形式）在其經處理之表面上。 經濟部智慧財產局員工消費合作社印製 發明簡述 本發明關於一種塑膠載片，其於表面經化學處理後可 被用於在其表面上固定蛋白質、肽或小分子化合物。本發 明之塑膠載片因此可用以製造生物晶片。該塑膠載片具有 至少一個凹槽，其中諸凹槽之深度可爲相同或不同，且一 般係在約0.01 mm至最局約〇_5 mm之範圍內 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐）~ — -5- 1275797 A7 B7 五、發明説明（3 ) 〇 · (請先閲讀背面之注意事項再填寫本頁) 本發明亦關於.由聚苯乙烯製成之塑膠載片，其於表面 經化學處理後可被用於在其經處理之表面上固定寡核苷酸 或聚核苷酸，故可用以製造D NA晶片。該塑膠載片具有 至少一個凹槽，其中諸凹槽之深度可爲相同或不同，且一 般係在約0.01 mm至最高約0.5 mm之範圍內 〇 圖_武之簡要說明 圖1示出本發明一較佳體系之塑膠載片的透視圖。 圖2 A示出本發明之經表面處理、且經C y 3 — SA / C y 5 - S A標記之塑膠載片的螢光影像，其中綠色螢 光點爲經C y 3 - S A標記之點，紅色螢光點爲經C y 5 —S A標記之點。 圖2 B示出先前技藝中之塑膠載片的螢光影像。 發明詳沭 經濟部智慧財產局員工消費合作社印製 本發明關於一種塑膠載片，其於表面經化學處理後可 被用於在其表面上固定蛋白質、肽或小分子化合物以製造 生物晶片，其中該塑膠載片具有至少一個凹槽，其中諸凹 槽之深度可爲相同或不同，且一般係在約〇 . 〇 1 mm 至最高約0 · 5 m m之範圍內。利用本發明之塑膠載片 所製造之生物晶片可被用作進行標的物篩選法（s h 〇 t g u η screening)時之平台，該標的物篩選法能以標的物導向之 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） _ 6 - 1275797 A7 B7 經濟部智慧財產局員工消費合作社印製 五、發明説明（4) 方式篩選出生物活性物質，而達到高效結果（high throughput ) ° 本發明塑膠載片之材料可爲同聚物或共聚物，其係由 1種或多種單體所製備，該單體係選自乙烯，鹵乙烯，丙 烯，鹵丙烯，丙烯酸酯，曱基丙烯酸酯，丁二烯，丙烯腈 ，原冰片烯與苯乙烯中，其中苯乙烯之聚合物係適宜的。 本發明塑膠載片之材料亦可爲聚碳酸酯。該塑膠載片之尺 寸係相當於微陣列器或雷射掃描器所慣用之載片尺寸。 使用本發明中具有特殊構造之塑膠載片的優點係在於 ，當與先前技藝中之平坦塑膠載片比較下，可看出所產生 之背景訊號（其在對生物晶片作分析時會產生干擾）較低 得多。 使用本發明之塑膠載片之另一優點係在於，可使用各 種不同之化學藥劑以處理或修飾塑膠載片之表面，使得不 僅是大分子化合物（諸如蛋白質和D N A )，亦包括小分 子化合物（諸如植物或草藥之代謝物）可固定於該塑膠載 片之表面上。此優點若對照慣用之玻璃載片僅能用於固定 大分子化合物（諸如蛋白質和D N A )之事實尤顯得重要 。再者，本發明之塑膠載片可易於模製成所欲之形狀，且 亦具有成本低之優異性。於本發明之較佳體系中，塑膠載 片係經模製以具有至少2個凹槽，使得於同一個塑膠載片 之凹槽中可同時進行至少2個不同之反應。每一個凹槽之 深度可爲相同或不同，且係介於約0 . 〇 1 m m至最高 約0 _ 5 m m之間。再者，每一個凹槽之相對兩邊可分 (請先閱讀背面之注意事項再填寫本頁) 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） 1275797 Α7 Β7 五、發明説明（5 ) 別模製欄桿以用於支撐玻璃蓋，其中該玻璃蓋係用於防止 載入至該凹槽內之溶液被蒸發或損失。 於製備表面經化學處理之塑膠載片中，係利用多官能 基醛及隨後浸沒於提供N Η 2基團之先驅物溶液中以預處 理上述之塑膠載片，使得經預處理之塑膠載片於其表面上 含有具活性之胺基。該提供Ν Η 2基團之先驅物可爲有機 物或無機物，且可選自Ν Η 4〇Η，一級胺，二級胺與叔 胺中，其中該一級胺，二級胺及叔胺之脂肪族及/或芳香 族部分可用於充作額外之間隔子。於該提供Ν Η 2基團之 先驅物中，直接提供自由Ν Η 2基團之Ν Η 4〇Η和一級 胺係適宜的。 進一步，令該經預處理之塑膠載片塗覆一層充作間隔 子之多官能基分子（例如，多官能基環氧化物）以製備本 發明之表面經化學處理之塑膠載片。於功能上，該多官能 基環氧化物之作用係連接固定於該表面經化學處理之塑膠 載片上的測試樣品（適宜地呈微陣列之型式）中所欲之成 份。該多官能基環氧化物之一端的活性環氧基係與該經預 處理之塑膠載片的表面上之胺基反應，而該多官能基環氧 化物之另一端的活性環氧基係吸附測試樣品中所欲之成份 或與其反應。詳言之，測試樣品中含有自由羥基，氫硫基 及/或胺基之成份能與該多官能基環氧化物之另一端的活 性環氧基反應以形成至少1個共價鍵，因而該成份能連接 至該表面經化學處理之塑膠載片上。該多官能基環氧化物 適宜地係含有6至2 4個碳原子之長化學鏈，使得所欲之 本紙張尺度適用中國國家標準（CNS ) Α4規格（210x297公釐） (請先閱讀背面之注意事項再填寫本頁) 衣· 訂 經濟部智慧財產局員工消費合作社印製 -8 - 1275797 A7 B7 五、發明説明（6) 成份不會直接地結合或吸附至該預處理之塑膠載片上。該 所欲之成份與表面經化學處理之塑膠載片之結合係牢固的 ，甚至經嚴厲之脫離沖洗後亦然。於本發明中，不僅是大 分子化合物（諸如蛋白質和多肽），亦包括小分子化合物 (諸如植物或草藥之代謝物）皆可以均一相或不均相之方 式固定於表面經化學處理之塑膠載片的表面上。 因此，製備表面經化學處理之塑膠載片包含下列之步 驟：取得具有凹槽之塑膠載片，利用多官能基醛及隨後浸 沒於提供N Η 2基團之先驅物溶液（適宜地係N Η 4〇Η 溶液）中以預處理該塑膠載片，及利用多官能基分子（適 宜地係多官能基環氧化物）以塗覆該經預處理之塑膠載片 之表面。 · 表面經化學處理之塑膠載片可用於製備生物晶片，其 中含有均一相或不均相之微陣列樣品斑點可固定於表面經 化學處理之塑膠載片的表面上。所得到之生物晶片可用於 以標的物導向之方式篩選出測試樣品斑點中所欲之成份。 上述之篩選方法可包含下列之步驟：將含有標記探針之溶 液載入上述之生物晶片中以進行化學反應（其中每一個凹 槽可覆蓋玻璃片以防止該含有標記探針之溶液的蒸發）， 及藉由利用裝置（例如雷射掃描器）以顯影並鑑定出能與 5亥標sH探針反應或結合之樣品斑點。供標記探針用之標記 物可爲染料或放射性物質。再者，用於雜交反應之探針可 爲基於已界定之分子機轉而呈均一相或不均相之已知標記 物，其係爲，例如，小分子化合物、競爭性配體或抗體， 本紙張尺度適用中國國家標準（CNS ) A4規格（210 X 297公釐） (請先閱讀背面之注意事項再填寫本頁) 衣· 經濟部智慧財產局員工消費合作社印製 -9 - 1275797 Α7 Β7 五、發明説明（7 ) 其能拮抗諸如經選擇之細胞、受體、酶或蛋白質。 (請先閲讀背面之注意事項再填寫本頁) 另一方面，本發明亦關於由聚苯乙儲材質製成的塑膠 載片，其於表面經化學處理後可被用於在其經處理之表面 上固定寡核苷酸或聚核苷酸，適宜地以微·陣列之方式，因 而可用於製造D N A晶片。該塑膠載片具有至少一個凹槽 ，其中諸凹槽之深度可爲相同或不同，且一般係在約 0.01 mm至最高約0.5 mm之範圍內。由本發 明之表面經處理之聚苯乙烯材質的塑膠載片所製造之 D N A晶片，可被用作偵測D N A用之平台，該平台可藉 由利用標記探針於雜交條件下偵測出於樣品斑點中是否存 有所欲之D N A。 經濟部智慧財產局員工消費合作社印製 本發明所使用之聚苯乙烯材質的塑膠載片之尺寸，亦 相當於微陣列器或雷射掃描器所慣用之載片尺寸。如前所 述，該聚苯乙烯材質之塑膠載片亦可經模製以適宜地具有 至少2個凹槽，使得於同一個塑膠載片之凹槽中可同時進 行至少2個不同之雜交反應。每一個凹槽之深度可爲相同 或不同，且係介於低於〇 · 01mm至高達〇 . 5mm之 間。再者，每一個凹槽之相對兩邊可分別模製欄桿以用於 支撐玻璃蓋，其中該玻璃蓋係用於防止載入至該凹槽內之 溶液被蒸發或損失。 對於修飾聚苯乙烯材質之塑膠載片的表面，係於較強 之鹼性條件下，利用試劑溶液塗覆聚苯乙烯材質之原始塑 膠載片，該試劑溶液包含不含有N Η 4 +基團之緩衝液且 該緩衝液含有提供正電荷之聚合物（例如聚離胺酸）。該 本紙張尺度適用中國國家標準（CNS ) Α4規格（210Χ297公釐） -10- 1275797 A7 B7 五、發明説明（8 ) 不含有N Η 4 +基團之緩衝液可爲碳酸鹽緩衝液、磷酸鹽 緩衝液或檸檬酸鹽緩衝液。上述較強之鹼性條件可爲介於 Ρ Η 9至1 1之間。製備本發明之表面經處理的聚苯乙烯 材質之塑膠載片的優點係在於僅需要單一步驟塗覆簡單之 試劑溶液。 上述所得到之D Ν Α晶片可藉由利用標記探針於雜交 條件下偵測出於測試樣品斑點中是否存有所欲之DNA。 該偵測方法可包含下述之步驟：將含有標記探針之溶液載 入上述之D Ν A晶片中以進行雜交反應（其中每一個凹槽 可覆蓋玻璃片以防止該含有標記探針之溶液的蒸發），及 藉由利用裝置（例如雷射掃描器）以顯影並鑑定出能與該 標記探針結合之樣品斑點。如前所述，供標記探針用之標 記物可爲染料或放射性物質。 於效果上，利用本發明之表面經處理的塑膠載片所製 造之生物晶片及D Ν A晶片皆比玻璃載片所製造之生物晶 片顯現出較弱得多之螢光背景。 本發明係以下述之實施例加以進一步之闡釋和說明， 但須明暸的是本發明之範圍並不以下述之實施例爲限。 實施例 利用螢光團標記蛋ή質或肽 依據製造商所建議之處理方法，利用 Fluori Link ™ Cy3 ™雙官能基反應染料（Amersham )及 FluoriLink ™ Cy5 ™雙官能基反應染料（Amersham )，分 本紙張尺度適用中國國家標準（CNS ) A4規格（2!〇><297公釐） (請先閲讀背面之注意事項再填寫本頁) 衣· 訂 經濟部智慧財產局員工消費合作社印製 -11 - 1275797 A7 B7 _ 五、發明説明（9 ) (請先閱讀背面之注意事項再填寫本頁) 別製備C y 3 - S A (亦即經C y 5標記之鏈抗生物素g 白（strepavidin，SA))及 C y 5 — S A (亦即經 C y 5 標記之鏈抗生物素蛋白）。使用在T B S T緩衝液（5 0 mMTris-HCl ，ρΗ 7·35，含有〇·15 M NaCl)中平衡之 Bio — Gel P 2 ( B i 0 一 Rad)管柱（0 . 5cmxl〇cm)進行膠體過濾 層析法，以移除未偶合之螢光殘基。估算最終濃度爲 0 · 5 m g / m 1 〇 塑膠載片之預處理及經塗覆塑膠載片之製備 經模製之塑膠載片及平坦之塑膠載片皆係由苯乙烯聚 合物製成。經模製之塑膠載片，係具有與微陣列器或雷射 掃描器所使用之慣用玻璃載片相當的尺寸，且係具有2個 凹槽，該凹槽之深度皆爲0 · 0 5 m m。平坦之塑膠載片 ，亦具有與微陣列器或雷射掃描器所使用之慣用玻璃載片 相當的尺寸。 經濟部智慧財產局員工消費合作社印製 經模製之塑膠載片及平坦之塑膠載片皆浸沒於0 · 4 %戊二醛之水溶液（p Η 5 . 0 )中4小時，隨後以水 加以沖洗，再於6 0 °C下浸入於3 Μ Ν Η 4〇Η水溶液 (Ρ Η 1 1 · 〇 )中4小時。於3 7 下，利用1 0 0 m Μ 1 ，. 4 一 丁二醇二縮水甘油醚（Ρ Η 1 1 · 〇 )隔 夜處理所生成之塑膠載片。所得之塑膠載片再以〇 . 1 Μ N a H C〇3水溶液（ρ Η 8 · 0 )沖洗一次，然後再 用二次蒸餾水沖洗四次。隨後可存放在4 °C之二次蒸餾水 本紙張尺度適用中國國家標準（CNS ) A4規格（210X 297公釐） -12- 1275797 A7 B7 五、發明説明（10) 中4〜1 6小時，或可省略此步驟。所得之載片於被使用 前，係在安全室內以室溫乾燥。 (請先閱讀背面之注意事項再填寫本頁)1275797 A7 B7 V. INSTRUCTION DESCRIPTION (J) FIELD OF THE INVENTION The present invention relates to plastic slides that can be used to make biochips. DETAILED DESCRIPTION OF THE INVENTION The present invention relates to a molded plastic slide having a special configuration which can be used to prepare a chemically treated plastic slide on which a biological sample of a microarray can be immobilized and spotted. Thus, a biochip is fabricated. BACKGROUND OF THE INVENTION Glass slides have been widely used as substrates for the manufacture of D N A wafers and protein wafers. However, the use of glass slides as substrates for biochips to date has revealed several disadvantages. First, the glass slide itself is fragile and requires careful handling during handling. The surface of the glass slide is not susceptible to chemical treatment in order to successfully immobilize biological small molecule compounds (such as plant or herbal metabolites) onto the treated surface for direct bonding. The glass slide also produces an undesirably high background signal that can interfere with subsequent analysis of the biofilm processed by the label. Therefore, this art requires the development of new materials or the development of materials that can be easily modified or modified to replace glass materials as substrates for biochips. Recently, some efforts have been made to develop biochips with plastic slides as substrates. The plastic slides used as the substrate of the wafer are not only less expensive to obtain than the glass slides, but also can be used by conventional injection molding. It is easily molded into the desired shape. w 0 0 0 / 5 5 6 2 7 discloses a biochip for detecting a target analyte, which comprises a paper size fixed on a non-fluorescent acrylic substrate, which is applicable to the Chinese National Standard (CNS) A4 specification ( 210 X 297 mm) K - n^— m —ϋΜ mu ·Γϋ m· mi -...- (Please read the notes on the back and fill out this page) Γ Γ Γ Γ Γ Γ Γ Γ Co-operative printing -4- 1275797 A7 ____ B7 ___ V. Description of invention (2) A list of biological binding ligands. W 0 0 0 / 3 6 1 4 5 discloses a method of manufacturing a biochip (please read the note on the back side and then fill out this page) method, which comprises grafting a biological probe onto a conductive polymer. EP 1 026 259 discloses a DNA wafer comprising a solid support and an oligonucleotide or polynucleotide immobilized on the solid support in the presence of a hydrophilic polymer. Although plastic materials have been used to make DNA wafers, the plastic slides used to date have been flat, that is, there are no grooves or protrusions on the upper surface, and they generate unwanted background signals. It can cause interference when analyzing biochips. Surface treated plastic slides have not been successfully used to immobilize (suitably in the form of microarrays) proteins, peptides or small molecule compounds onto their treated surfaces, particularly in the protein, peptide or small molecule The compound is unmodified. In addition, the prior art does not teach or suggest that the surface of a plastic slide of polystyrene can be treated in a one-step manner by using a simple reagent' to enable the oligonucleotide or polynucleotide to be immobilized (suitably In the form of a microarray) on its treated surface. The invention relates to a plastic slide which can be used to immobilize a protein, peptide or small molecule compound on its surface after chemical treatment on the surface. The plastic slide of the present invention can therefore be used to make a biochip. The plastic carrier has at least one groove, wherein the depths of the grooves may be the same or different, and generally range from about 0.01 mm to a maximum of about 〇5 mm, and the paper size is applicable to the Chinese National Standard (CNS). A4 size (210X297 mm)~ — -5- 1275797 A7 B7 V. Description of invention (3) 〇· (Please read the note on the back and fill out this page) The invention also relates to plastic made of polystyrene A slide, which can be used to immobilize an oligonucleotide or a polynucleotide on its treated surface after chemical treatment on the surface, can be used to make a DAA wafer. The plastic carrier has at least one groove, wherein the depths of the grooves may be the same or different, and generally range from about 0.01 mm to up to about 0.5 mm. FIG. 1 shows a first embodiment of the present invention. A perspective view of a preferred system of plastic slides. Figure 2A shows a fluorescent image of a surface-treated, Cy 3 - SA / C y 5 - SA-labeled plastic slide of the present invention, wherein the green fluorescent spot is marked by Cy 3 - SA The red fluorescent spot is the point marked by C y 5 —SA. Figure 2B shows a fluorescent image of a prior art plastic slide. The invention relates to a plastic slide which can be used for chemically treating a surface to immobilize a protein, peptide or small molecule compound on its surface to produce a biochip, wherein The plastic carrier has at least one groove, wherein the depths of the grooves may be the same or different, and generally range from about 〇1 mm to a maximum of about 0.5 mm. The biochip manufactured by using the plastic slide of the present invention can be used as a platform for performing the screening method, which can be applied to the national standard of the standard-oriented paper scale. (CNS) A4 size (210X297 mm) _ 6 - 1275797 A7 B7 Ministry of Economic Affairs Intellectual Property Bureau employee consumption cooperative printing 5, invention description (4) Way to screen out biologically active substances, and achieve high efficiency (high throughput) The material of the inventive plastic slide may be a homopolymer or a copolymer prepared from one or more monomers selected from the group consisting of ethylene, vinyl halide, propylene, halopropylene, acrylate, mercapto acrylate. Among the butadiene, acrylonitrile, norbornene and styrene, the polymer of styrene is suitable. The material of the plastic slide of the present invention may also be polycarbonate. The size of the plastic carrier is equivalent to the size of the slide used by the microarray or laser scanner. The advantage of using a specially constructed plastic slide of the present invention is that when compared to a flat plastic slide of the prior art, the resulting background signal (which interferes with the analysis of the biochip) can be seen. Much lower. Another advantage of using the plastic slide of the present invention is that a variety of different chemicals can be used to treat or modify the surface of the plastic slide so that it is not only macromolecular compounds (such as proteins and DNA) but also small molecule compounds ( A metabolite such as a plant or herb can be immobilized on the surface of the plastic slide. This advantage is particularly important in comparison to the fact that conventional glass slides can only be used to immobilize macromolecular compounds such as proteins and D N A . Further, the plastic slide of the present invention can be easily molded into a desired shape, and also has a low cost superiority. In a preferred embodiment of the invention, the plastic carrier is molded to have at least two grooves such that at least two different reactions can occur simultaneously in the grooves of the same plastic carrier. The depth of each groove may be the same or different and is between about 0. 〇 1 m m and a maximum of about 0 _ 5 m m. Furthermore, the opposite sides of each groove can be divided (please read the note on the back and fill in the page first). The paper size applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) 1275797 Α7 Β7 V. Invention description ( 5) Do not mold the railing for supporting the glass cover, wherein the glass cover is used to prevent the solution loaded into the groove from being evaporated or lost. In preparing a chemically treated plastic slide, the pre-treated plastic slide is pretreated by using a polyfunctional aldehyde and subsequently immersed in a precursor solution providing a N Η 2 group to pretreat the plastic slide. Containing an active amine group on its surface. The precursor providing the Ν 2 group may be organic or inorganic, and may be selected from the group consisting of Ν 4Η, a primary amine, a secondary amine and a tertiary amine, wherein the primary amine, the secondary amine and the tertiary amine are fat. The family and/or aromatic moiety can be used as an additional spacer. In the precursor of the group providing the Ν 2 group, the Ν 4 〇Η and the primary amine which are directly available for the free Ν 2 group are suitable. Further, the pretreated plastic carrier is coated with a layer of polyfunctional molecules (e.g., polyfunctional epoxide) to form a surface chemically treated plastic carrier of the present invention. Functionally, the polyfunctional epoxide acts as a desired component in a test sample (preferably in the form of a microarray) immobilized on the chemically treated plastic slide. The reactive epoxy group at one end of the polyfunctional epoxide reacts with an amine group on the surface of the pretreated plastic carrier, and the reactive epoxy group at the other end of the polyfunctional epoxide is adsorbed. Test or react with the desired ingredients in the sample. In particular, the test sample contains a free hydroxyl group, a thiol group and/or an amine group component capable of reacting with the active epoxy group at the other end of the polyfunctional epoxide to form at least one covalent bond, thus The ingredients can be attached to the chemically treated plastic slide on the surface. The polyfunctional epoxide is suitably a long chemical chain of 6 to 24 carbon atoms, so that the desired paper size is applicable to the Chinese National Standard (CNS) Α 4 specification (210 x 297 mm) (please read the back) Precautions and then fill out this page) Clothing · Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -8 - 1275797 A7 B7 V. Invention Description (6) The ingredients are not directly bonded or adsorbed onto the pretreated plastic slide. The combination of the desired ingredients and the chemically treated plastic slides is strong, even after severe rinsing. In the present invention, not only macromolecular compounds (such as proteins and polypeptides) but also small molecule compounds (such as plant or herbal metabolites) can be immobilized on the surface of chemically treated plastics in a uniform or heterogeneous manner. On the surface of the piece. Thus, the preparation of a chemically treated plastic carrier comprises the steps of obtaining a plastic carrier having a recess, utilizing a polyfunctional aldehyde and subsequently immersing in a precursor solution providing a N Η 2 group (suitably N Η The plastic slide is pretreated in a solution to coat the surface of the pretreated plastic slide with a polyfunctional molecule, suitably a polyfunctional epoxide. • A chemically treated plastic slide can be used to prepare a biochip with a uniform or heterogeneous microarray sample spot that can be attached to the surface of a chemically treated plastic slide. The resulting biochip can be used to screen out the desired components of the test sample spot in a target-oriented manner. The screening method described above may comprise the steps of loading a solution containing the labeled probe into the biochip described above for chemical reaction (where each groove covers the glass sheet to prevent evaporation of the solution containing the labeled probe) And by using a device (such as a laser scanner) to develop and identify sample spots that can react or bind with the 5H sH probe. The label for the labeled probe can be a dye or a radioactive material. Furthermore, the probe for the hybridization reaction may be a known label which is a homogeneous phase or a heterogeneous phase based on a defined molecular machine, which is, for example, a small molecule compound, a competitive ligand or an antibody, This paper scale is applicable to China National Standard (CNS) A4 specification (210 X 297 mm) (please read the note on the back and fill out this page). Clothing · Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -9 - 1275797 Α7 Β7 V. INSTRUCTIONS (7) It antagonizes, for example, selected cells, receptors, enzymes or proteins. (Please read the precautions on the back and then fill out this page.) On the other hand, the present invention also relates to a plastic slide made of polystyrene storage material which can be used for chemical treatment after being chemically treated on the surface. Oligonucleotides or polynucleotides are immobilized on the surface, suitably in the form of microarrays, and thus can be used to fabricate DNA wafers. The plastic carrier has at least one recess, wherein the depths of the grooves may be the same or different, and are generally in the range of from about 0.01 mm up to about 0.5 mm. The DNA wafer manufactured by the surface treated polystyrene plastic slide of the present invention can be used as a platform for detecting DNA, and the platform can be detected by using a labeled probe under hybridization conditions. Whether there is a DNA in the spot. Printed by the Intellectual Property Office of the Ministry of Economic Affairs, the Consumers' Cooperatives. The size of the plastic slides made of polystyrene used in the present invention is also equivalent to the size of the slides conventionally used in microarrays or laser scanners. As mentioned above, the plastic slide of polystyrene material can also be molded to suitably have at least 2 grooves, so that at least two different hybridization reactions can be simultaneously performed in the grooves of the same plastic slide. . The depth of each groove may be the same or different and is between 〇 · 01mm and up to 〇 5 mm. Further, the opposite sides of each of the grooves may be separately molded with railings for supporting the glass cover, wherein the glass cover is for preventing evaporation or loss of the solution loaded into the grooves. For the surface of the plastic slide modified with polystyrene material, under the strong alkaline condition, the original plastic slide of polystyrene material is coated with the reagent solution, and the reagent solution contains no N Η 4 + group. The buffer and the buffer contains a polymer that provides a positive charge (eg, polyaminic acid). The paper size is applicable to the Chinese National Standard (CNS) Α4 specification (210Χ297 mm) -10- 1275797 A7 B7 5. Inventive Note (8) The buffer containing no N Η 4 + group can be carbonate buffer, phosphoric acid Salt buffer or citrate buffer. The above strong alkaline conditions may be between Ρ Η 9 and 11. The advantage of preparing a surface treated polystyrene plastic slide of the present invention is that only a single step of coating a simple reagent solution is required. The D Ν Α wafer obtained above can be used to detect whether or not the desired DNA is present in the test sample spot by using a labeled probe under hybridization conditions. The detecting method may comprise the steps of: loading a solution containing the labeled probe into the above D Ν A wafer for hybridization reaction (wherein each groove covers the glass sheet to prevent the solution containing the labeled probe) Evaporation), and by using a device (such as a laser scanner) to develop and identify sample spots that can bind to the labeled probe. As previously mentioned, the label for the labeled probe can be a dye or a radioactive material. In effect, the biochip and D Ν A wafers produced using the surface treated plastic slide of the present invention exhibit a much weaker fluorescent background than the biochip produced by the glass slide. The invention is further illustrated and described by the following examples, but it should be understood that the scope of the invention is not limited EXAMPLES Fluori LinkTM Cy3TM Bifunctional Reactive Dyes (Amersham) and FluoriLinkTM Cy5TM Bifunctional Reactive Dyes (Amersham) were used to label egg tarts or peptides using fluorophores according to the manufacturer's recommended treatment. This paper size applies to the Chinese National Standard (CNS) A4 specification (2!〇><297 mm) (please read the notes on the back and then fill out this page) Clothing and Customs Ministry of Economic Affairs Intellectual Property Bureau Staff Consumer Cooperative Printed -11 - 1275797 A7 B7 _ V. INSTRUCTIONS (9) (Please read the note on the back and then fill out this page) Do not prepare C y 3 - SA (ie, the chain labeled with avidin g by C y 5 ( Strepavidin, SA)) and C y 5 — SA (ie, Cy 5 labeled streptavidin). Colloid using a Bio-Gel P 2 (B i 0 -Rad) column (0.5 cmxl〇cm) equilibrated in TBST buffer (50 mMTris-HCl, ρΗ 7.35, containing 〇·15 M NaCl) Filtration chromatography was performed to remove uncoupled fluorescent residues. Estimated final concentration of 0 · 5 m g / m 1 预处理 Pretreatment of plastic slides and preparation of coated plastic slides Molded plastic slides and flat plastic slides are made of styrene polymer. The molded plastic slide has a size comparable to that of a conventional glass slide used in a microarray or laser scanner, and has two grooves having a depth of 0 · 0 5 mm . Flat plastic slides also have dimensions comparable to conventional glass slides used in microarray or laser scanners. The plastic slides and flat plastic slides printed by the Intellectual Property Office of the Intellectual Property Office of the Ministry of Economic Affairs were immersed in an aqueous solution of 0. 4 % glutaraldehyde (p Η 5.0) for 4 hours, followed by water. Rinse and immerse in a 3 Μ Η 4 〇Η aqueous solution (Ρ Η 1 1 · 〇) at 60 ° C for 4 hours. The plastic slides produced were treated overnight with 1 0 0 m Μ 1 , . 4 - butanediol diglycidyl ether (Ρ Η 1 1 · 〇) at 37 °C. The resulting plastic slide was rinsed once with a solution of 〇 1 Μ N a H C〇3 (ρ Η 8 · 0) and then rinsed four times with twice distilled water. The second distilled water can be stored at 4 °C. The paper size is applicable to the Chinese National Standard (CNS) A4 specification (210X 297 mm) -12- 1275797 A7 B7 5. The invention description (10) is 4 to 1 6 hours, or This step can be omitted. The resulting slides were dried at room temperature in a safe room before being used. (Please read the notes on the back and fill out this page)
1 動_ 配置 Cy3 — SA 菸 Cv5 — SA 經濟部智慧財產局員工消費合作社印製 利用配備有0 · 4mm固體針管之自動裝置 MicroGrid I I (購自 BioRobotics )，將經稀釋之 C y 3 - S A及C y 5 - S A配置於如上述之經模製塑膠 載片與平坦塑膠載片上之特定位置。使用3 〇% ( 1 0〜 60%) DMS〇/〇 . 1M碳酸鹽緩衝液，其具有pH 9 . 5 ( p Η 7 . 5〜1 1 ) ，2倍順序地稀釋C y 3 — SA 及 Cy5 — SA 蛋白質，使 Cy3 — SA 及 Cy5 — SA蛋白質於各稀釋液中之濃度爲2 0 // g/m 1至3 9 n g /m 1 。該固體針管係用於投遞含有蛋白質化合物之 溶液並形成斑點，因此於配置每一個樣品前，利用二次蒸 餾水沖洗該固體針管2秒，再利用7 0 % E t〇Η沖洗 該固體針管2秒，隨後置於熱空氣流中乾燥2秒。完成配 置樣品後，該經模製塑膠載片與平坦塑膠載片皆於室溫下 被培育2小時，隨後將其浸沒於1 Μ乙醇胺中。隨後利用 T B S 丁 緩衝液（5 0 m Μ T r i s · H C 1 ， 〇.15M N a C 1 ^ 0 . 0 5 % Tween 2 0 )沖 洗所得到之生物晶片3次，再以二次蒸餾水輕洗3次，進 而置於3 7 °C下乾燥5分鐘。接著利用GenePix 4000A載 片掃描器（Axon Instruments)掃描’及利用 GenePix 3.0 軟體加以分析所得之影像。結果顯示於圖2 A與2 B中， 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -13- 1275797 A7 _____B7 _ 五、發明説明（1彳） 其中圖2 A與2 B所示之經配置樣品係各依順序地由第1 至1 0欄（由左至右）2倍地稀釋。 每一樣品在橫列組中之濃度由左至右爲如下，其中各 橫列組中所包含之第二列係爲重複第一列之操作所得： 橫列組（a )與（b ):1 Move_Configure Cy3 — SA Tobacco Cv5 — SA Ministry of Economic Affairs Intellectual Property Office Staff Consumer Cooperative Printed using the MicroGrid II (purchased from BioRobotics) equipped with a 0·4mm solid needle, the diluted C y 3 - SA and C y 5 - SA is disposed at a specific location on the molded plastic carrier and the flat plastic carrier as described above. Use 3 〇% (1 0~60%) DMS〇/〇. 1M carbonate buffer with pH 9.5 (p Η 7 . 5~1 1 ), 2 times sequentially diluted C y 3 — SA and Cy5 - SA protein, the concentration of Cy3-SA and Cy5-S protein in each dilution is 20 @ g / m 1 to 3 9 ng / m 1 . The solid needle tube is used for delivering a solution containing a protein compound and forming a spot, so the solid needle tube is rinsed with double distilled water for 2 seconds before each sample is configured, and the solid needle tube is rinsed with 70% E t〇Η for 2 seconds. It was then dried in a stream of hot air for 2 seconds. After the configuration of the sample was completed, the molded plastic slide and the flat plastic slide were incubated at room temperature for 2 hours, and then immersed in 1 Μ ethanolamine. The obtained biochip was then washed 3 times with TBS buffer (50 m Μ T ris · HC 1 , 〇.15M N a C 1 ^ 0 . 0 5 % Tween 2 0 ), and then lightly washed with double distilled water. 3 times, and then dried at 3 7 ° C for 5 minutes. The images were then scanned using the GenePix 4000A slide scanner (Axon Instruments) and analyzed using GenePix 3.0 software. The results are shown in Fig. 2 A and 2 B. The paper scale is applicable to the Chinese National Standard (CNS) A4 specification (210X297 mm) -13- 1275797 A7 _____B7 _ V. Invention description (1彳) where 2 and 2 B The configured samples shown are each diluted 2 times sequentially from columns 1 to 10 (left to right). The concentration of each sample in the row group is from left to right as follows, wherein the second column included in each row group is obtained by repeating the operation of the first column: row groups (a) and (b):
Cy5-SA: 20, 1 0, 5, 2.5, 1.25, 0.625, 0.3 1 2, 0.1 56, 0.078, 0·039 pg/ml 橫列組（c )與（d ):Cy5-SA: 20, 1 0, 5, 2.5, 1.25, 0.625, 0.3 1 2, 0.1 56, 0.078, 0·039 pg/ml Alignment groups (c) and (d):
Cy3_SA: 20，10，5，2.5，1.25, 0.625, 0.3 1 2, 0.1 56, 0.078， 0.039 pg/ml (請先閱讀背面之注意事項再填寫本頁) 經濟部智慧財產局員工消費合作社印製 本紙張尺度適用中國國家標準（CNS ) A4規格（210X297公釐） -14-Cy3_SA: 20,10,5,2.5,1.25, 0.625, 0.3 1 2, 0.1 56, 0.078, 0.039 pg/ml (Please read the note on the back and fill out this page) Printed by the Consumer Intellectual Property Office of the Ministry of Economic Affairs This paper scale applies to the Chinese National Standard (CNS) A4 specification (210X297 mm) -14-