本研究目的在於自行合成甲基安非他命(methamphetamine; Meth)與蛋白質之生物共軛物(bioconjugates)來腹腔免疫ICR小鼠，以生產Meth 之多株抗體(P olyclonal antibodies against methamphetamine : anti-Meth PAbs)。由於Meth之分子量很小(約149.233 g/mol)不易在小鼠體內引起免疫反應，故本研究選用牛血清白蛋白(bovine serum albumin；BSA)以及絲膠蛋白(Sericin ; Ser)，分別與Meth 形成生物共軛物，藉由大分子蛋白誘導小鼠體內產生Meth 之抗體。多株抗體製作過程為將高度免疫hyperimmune)中之ICR小鼠，注射0.5 mL pristane 後，再間隔兩週注射106l個NS-1 骨髓癌細胞。待小鼠腹腔漲大至行動困難時，收集ICR 小鼠所產生之腹水及血清，利用HitrapTM rProtein A column 來進行多株抗體之初步純化，並進一步測定anti-Meth PAbs 之效價(titer)以及進行蛋白質定量工作。本實驗工作首先成功合成出藥物與蛋白質之生物共軛物，其中Meth 與BSA 之生物共軛物(Meth-BSA)當作免疫原(immunogen)進行小鼠免疫工作；另外，Meth 與Ser 之生物共軛物(Meth-ser)當作酵素連結免疫吸附分析法(Enzyme-linked immunosorbent assay ; ELISA)檢測系統中所用之抗原(antige)，以避免測到腹水中anti-BSA 抗體效價，而造成誤差• 最後生產並初步純化5.68mg /mL多株抗體(含anti-Meth and anti-BSA PAbs)，並加以冷凍乾燥保存。 The aim of this study was to synthesize methaphetamine (Meth) - protein bioconjugates and produce polyclonal antibodies (PAbs) against Meth (anti-Meth PAbs ) . ICR mice were intraperitoneally immunized with Meth-protein bioconjugates every two weeks. Because the molecular weight of Meth is very small , it is difficult to induce production of mice antibodies The Meth molecules were conjugated with bovine serum albumin ( BSA ) and sericin (Ser), respectively. Hyperimmune ICR mice produced anti-Meth PAbs after injected with 0.5 mL pristane, and injected with NS-1 rnyeloma cells two weeks later. Ascites and serum were collected from ICR mice, but before the rnice had difficulty moving, and anti-Moth PAbs was Purified by using HitrapTM rProtein A column. In this study, the drug-proteins bioconjugates had been successfully produced. Meth-BSA bioconjugates were as the immunogen for mice immunization and Meth-Ser bioconjugates were as the antigen for Enzyme-Link Immunosorbent Assay (ELISA). Finally the production and primary purification of a anti-Meth PAbs were performed The concentration of PAbs obtained was 5.68 mg / mL. Then, the anti-Meth PAbs were stored by freeze-dry method .
