抗人血管生长素噬菌体基因工程单链抗体的初步制备

PREPARATIONOFTHEPHAGEGENETICENGINEERING ANTIBODYOFANTI-HUMAN ANGIOGENIN

采用ＰＣＲ等分子克隆实验手段从小鼠杂交瘤细胞中获取抗人血管生长素抗体可变区重、轻链基因，以ＤＮＡ接头连接后与噬菌粒ｐＣＡＮＴＡＢ５Ｅ重组转化大肠杆菌形成抗体可变区基因库。通过噬菌体表面表达技术把抗体可变区表达在噬菌体表面。

Thephage display antibody ofanti-hum an angiogenin w asanew and high effective antibody prepared by using the techniqueofsurfacedisplay phage antibody library.By using m olecularbiologicalm ethodssuch asPCRet al,im m unoglobulin variableregion geneswere am plified from angiogenin m ousehybridom as.Theheavy and lightchain DNA productsw ere assem bled into a single gene by using a DNA linker.The anti-angiogenin single DNA fragm ent w as ligated into a phage vector pCANTAB5E,and the recom binantvectorw assubsequently transform ed into E.coli TG1 cells forconstruction ofphage display librariesofanti-angiogenin.By phage rescue、affinity selection、reinfection and screening m ethods,the anti-angiogenin phage geneticengineering single chain antibody ofdefined specificity and affinity w as obtained.