Völkel Tina, Müller Rolf, Kontermann Roland E
Institut für Molekularbiologie und Tumorforschung (IMT), Philipps-Universität, Emil-Mannkopff-Str. 2, 35033 Marburg, Germany.
Biochem Biophys Res Commun. 2004 Apr 30;317(2):515-21. doi: 10.1016/j.bbrc.2004.03.074.
We have isolated single-chain Fv fragments directed against human endothelial cells from a novel fully synthetic human scFv library (scFv 479). This library was constructed using the variable germline segments DP47 and DPkappa9 as scaffolds. Complementarity determining regions 3 (CDR) of the variable heavy and light chain were introduced with a length of 9 amino acid residues. In total, 16 amino acid positions of all six CDRs exposed in the antigen-binding site were randomized and the library was produced from synthetic oligonucleotides encoding the entire scFv fragment. From this library endothelial-specific scFv fragments were either selected using the recombinant extracellular domain of human endoglin (CD105) or by cell selections with human dermal microvascular endothelial cells (HDMEC). These scFv fragments might be useful for the generation of vascular or tumor targeting agents in cancer therapy.
我们从一个新型的全合成人单链抗体片段（scFv）文库（scFv 479）中分离出了针对人内皮细胞的单链抗体片段。该文库是以可变种系片段DP47和DPkappa9为支架构建的。重链和轻链可变区的互补决定区3（CDR）引入了长度为9个氨基酸残基。总共，对抗原结合位点中暴露的所有六个CDR的16个氨基酸位置进行了随机化处理，并且该文库由编码整个单链抗体片段的合成寡核苷酸产生。从该文库中，使用人内皮糖蛋白（CD105）的重组胞外结构域或通过用人真皮微血管内皮细胞（HDMEC）进行细胞筛选来选择内皮细胞特异性单链抗体片段。这些单链抗体片段可能有助于在癌症治疗中生成血管或肿瘤靶向剂。
