本研究目的在使用 C-反應蛋白 (C-reactive protein, CRP) 腹腔免疫 ICR 小鼠，來生產 C-反應蛋白之多株抗體 (C-reactive protein polyclonal antibodies, anti-CRP)。將高度免疫 (hyperimmune) 之 ICR 小鼠，注射 0.5 mL pristine 後，再間隔兩週注射 106 個 NS-1 骨髓癌細胞。收集 ICR 小鼠所產生之腹水及血清，利用 HitrapTM rProtein A column 來純化多株抗體，並進一步分析已純化 anti-CRP 之抗體效價及其專一性。本研究以間接型酵素連結免疫分析法 (enzyme-linked immunosorbent assay, ELISA) 測定 anti-CRP 之效價。實驗結果顯示，多株抗體 anti-CRP 的效價為 1：3,125。可測得 C-反應蛋白最低濃度為 5 ng/mL，CRP 濃度範圍由 5 ng/mL 至 100 μg/mL 與波長 405nm 之吸光值呈良好線性關係，檢測濃度範圍可謂相當廣。 The aim of this study was to produce polyclonal antibodies against C-reactive protein (anti-CRP). ICR mice was immunized intraperitoneally with C-reactive protein (CRP) for four to five times every four weeks. Hyperimmune ICR mouse produced anti-CRP after injecting with 0.5 mL pristane, and injected with NS-1 myeloma cells two weeks later. Ascites and serum were collected from ICR mice, and anti-CRP was purified by using HitrapTM rProtein A column, and determination of the titer and specificity of anti-CRP using indirect enzyme-linked immunosorbent assay (ELISA). The results showed the titer of anti-CRP was 1:3,125, and low cross-reaction. A linear relationship existed between absorbance at 405 nm (A405) and CRP concentrations in the range from 5 ng/mL to 100 μg/mL. The detection range was very wide.
