论文部分内容阅读
目的：制备抗人肝癌单抗（ ＭｃＡｂ） ＨＡｂ１８ Ｆ（ａｂ’）２ 片段。方法 ：用二硫苏糖醇（ＤＴＴ） 激活木瓜蛋白酶，ｐＨ５ ．５ 环境下消化ＨＡｂ１８ ＭｃＡｂ（ＩｇＧ１） ，然后用快速蛋白液相色谱（ＦＰＬＣ） ＤＥＡＥ－ Ｓｅｐｈａｒｏｓｅ － ＦＦ 柱（２ｃｍ ×１８ｃｍ）纯化。对所获Ｆ（ａｂ’）２ 段的纯度、免疫活性、热原质、细菌、产率等方面进行质量检测。结果：在酶∶抗体为１∶２０ ，消化ＨＡｂ１８ ２ ｈ ，９５ ％ 以上ＩｇＧ 裂解为Ｆ（ａｂ’）２ 段，纯化后Ｆ（ａｂ’）２ 段的纯度为＞ ９６ ．１ ％ ，纯化周期６０ ｍｉｎ ， 一次制备量可达５００ ｍｇ ，产率为５３ ％ ，免疫活性１∶８０００ ，细菌、热原检测结果为阴性。结论：该法是一种快速、大量制备高产率、高质量ＭｃＡｂ ＩｇＧ１ 亚类Ｆ（ａｂ’）２ 段的有效方法。
Objective: To prepare anti-human hepatoma McAb HAb18 F (ab ’) 2 fragment. Methods: Papain was activated with dithiothreitol (DTT), pH5. HAb18 McAb (IgG1) was digested with EcoRI, 5, and then purified using a Fast Protein Liquid Chromatography (FPLC) DEAE-Sepharose-FF column (2 cm x 18 cm). The purity, immunocompetence, pyrogen, bacteria, yield and other aspects of the obtained F (ab ’) 2 were tested. Results: The fragment of F (ab ’) 2 was cleaved to> F (ab’) 2 after enzyme digestion with HAb18 at 1:20 and digestion of HAb18 for more than 95%. 1%, the purification cycle was 60 min, the initial preparation amount was 500 mg, the yield was 53%, the immunogenicity was 1: 8000, and the result of bacteria and pyrogen test was negative. Conclusion: This method is an efficient and rapid method for mass production of McAb IgG1 subclass F (ab ’) 2 with high yield and high quality.