C-反應蛋白多株抗體之製備與純化

Production and Purification of Polyclonal Antibodies against C-reactive Protein

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0.5mL 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5ng/mL，CRP濃度範圍由5ng/mL至100μg/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 0.5mL 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 Hitrap(Superscript TM) 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 405nm (A405) and CRP concentrations in the range from 5ng/mL to 100μg/mL. The detection range was very wide.