碳六十固定化抗體感測器的研製與應用

Abstract

本研究在研製一抗體的壓電晶體感測器，將塗佈碳六十的石英晶片作為研究碳六十與Anti-IgG抗體間的反應，以及運用此固定化之石英晶體感測器，偵測溶液中IgG抗體分子存在。結果顯示，碳六十能和Anti-IgG抗體結合，呈現出不可逆的化學吸附現象；碳六十與Anti-IgG抗體置於反應瓶反應，反應一天之後，發現有褐色的生成物，將此生成物經IR光譜的鑑定。並將其圖譜與碳六十光譜及抗體光譜做比較，得知產物是由碳六十及Anti-IgG抗體結合而成。 Anti-IgG抗體與碳六十的反應的濃度及碳六十的塗佈量亦做探討。而接上碳六十的抗體石英晶片，亦能偵測出IgG抗體分子，證實碳六十與抗體結合後依然有活性。碳六十及Anti-IgG抗體石英晶片於4℃的保存條件下，可以達到七天的保存期限。碳六十的塗佈飽和量為1.3mg。而Anti-IgG抗體與IgG抗體的反應條件受到pH值、溫度、濃度的影響。研究顯示其最適pH值於6.7，最適溫度為30℃。而此探針的偵測極限可到達3.25×10-4mg/mL。此碳六十/Anti-IgG抗體石英晶體感測器經過重複使用七次，仍然不錯的再現性。 本研究並對有機物質存在下，以石英晶體感測器感測Anti-IgG抗體和IgG抗體的反應，觀察有機物質對反應的影響。甲醇、乙醇、丙醇都做探討，發現其對本實驗有干擾。而苯甲酸、氨、丙酮、丙醛、苯甲醛的干擾也做探討。溶液中的鉛離子及銅離子的影響比較，銅離子影響本實驗較大。對於血液中的干擾物質如尿素(urea)、尿酸(uric acid)、維他命C(ascorbic acid)、半胱胺酸(cystein)、酪胺酸(tyrosine)亦做探討，發現其對於本實驗的影響並不大。鈉離子、鉀離子、鈣離子在10-3M濃度時，亦不會對本實驗造成干擾。 本研究亦利用石英壓電晶體感測器探討麥麩蛋白抗體及麥麩蛋白的作用。結果顯示固定上碳六十石英晶片的麥麩蛋白依然有活性。血紅蛋白及血紅蛋白抗體的亦做探討，結果顯示固定上碳六十石英晶片的血紅蛋白依然有活性，可以偵測到血紅蛋白抗體。Fullerene C60/anti-human IgG was synthesized and applied in a piezoelectric crystal immunosensor for human IgG .The C60 /anti-human IgG coated piezoelectric quartz crystal sensor was employed to study the interaction between anti-human IgG and human IgG. The immobilized C60/anti-human IgG was identified with IR spectrography. The antibody(anti-human IgG)immobilization on C60 was studied with a C60-coated piezoelectric quartz crystal detection system. The antibody immobilization was influenced by C60 coating load and IgG concentration. The immobilized antibody was still actived. The life time of immobilized C60/anti-human IgG was 85% after 7 days. The interaction of anti-human IgG and human IgG was influenced by pH, reaction temperature, and concentration. Optimum pH of immobilized C60/anti-human IgG at 6.7 and optimum temperature at 30℃ were found. The piezoelectric immuno-sensor for human IgG also has exhibited good sensitivity and good reproducibility for reusage. The detection limit of the sensor was 3.25´10-4mg/mL for human IgG. The interaction of anti-human IgG and human IgG affinity in organic solvents such as methanol, ketone, benzonic acid,ammonia was also studied. The interference of ions and other compounds in blood were also investiged and discussed. Various metal ions such as Na+, K+, Ca2+ in aqueous solutions showed nearly no interference to the detection of human IgG with the piezoelectric immunosensor. The piezoelectric quartz crystal sensor was also employed to study the interaction between anti-gliadin and gliadin. The immobilized gliadin was still actived. The interaction between Hemoglobin and anti-hemoglobin was also studied by piezoelectric quartz crystal sensor.