抗HRP单克隆抗体及PAP复合物活性影响因素的研究

A Study on the Factors to Affect the Activities of McAb against HRP and McPAP

作者制备了大鼠抗辣根过氧化物酶单克隆抗体及其与辣根过氧化物酶的复合物,观察了温度(室温、4℃、-20℃、-70℃)、pH值(pH=3,pH=11),蛋白稳定剂(1％BSA,50％中性甘油)及保存时间对抗体活性的影响。以ELISA法及微孔免疫细胞化学法检测抗体活性。结果表明;含此抗体的腹水及培养上清在室温中放置2周,4℃、-20℃、-70℃中放置10个月,抗体活性未见明显改变,加与不加蛋白稳定剂无明显差异。此培养上清在酸性环境(pH=3)中24h,活性严重受损,而其中含1％BSA及腹水者可耐受24h,活性无明显变化。此抗体与酶的复合物在4℃、-20℃、-70℃保存1年,仍保持良好的染色效果,与开始相比无显著性差异。

Rat monoclonal antibodies (McAb) against horseradish peroxidase (HRP) and HRP-anti-HRP McAb (McPAP) complex were made. It was observed that different temperatures (room temperature, 4℃, -20℃, -70℃), pH (pH = 3, pH=11), stabilizers (1% BSA, 50% neutral glycerine) and storage life affected the activities of McAb and McPAP, which were appraised by ELISA and microwell immunocytochemistry methods. When the ascitic fluid and culture supernatant of monoclonal antibodies were stored at room temperature for two weeks and temperature of 4℃,-20℃,-70℃, for ten months, the activities did not change evidently, whether the stabilizer was added or not. On the other hand, if the McAb supernatant was put under pH = 3 with Tris-HC1 buffer for 24 hours the activities of McAb decreased apparently. But the supernatant with 1% BSA of McAb was influenced less slightly by pH = 3 than that without 1% BSA. The facts are that whether supernatant is added with 1 % BSA and ascites or not, it has some defence role to the McAb under the ascitic condition (pH = 3). The activities of McPAP do not change at all, when McPAP is kept at the temperature of 4 ℃, - 20℃, - 70℃, for one year. Therefore, this McAb and McPAP are useful and satisfying reagents for the research and application in clinical work.