An approach for preventing recombination-deletion of the 40-50 anti-digoxin antibody V(H) gene from the phage display vector pComb3.
An approach for preventing recombination-deletion of the 40-50 anti-digoxin antibody V(H) gene from the phage display vector pComb3.
复制标题
一种防止噬菌体展示载体 pComb3 中 40-50 抗地高辛抗体 V(H) 基因重组删除的方法。
DOI：
10.1016/s0378-1119(99)00462-x
复制
发表时间：
2000
期刊：
影响因子：
3.5
通讯作者：
Margolies,MN
中科院分区：
文献类型：
--
作者：
Kim,SH;Titlow,CC;Margolies,MN
噬菌体展示已广泛应用于抗体 (Ab) 工程中。然而，有时噬菌体展示载体会表现出免疫球蛋白 (Ig) 基因的缺失。作为避免克隆到 pComb3 载体中的鼠抗地高辛 Fab 40-50 重组缺失的方法，通过用基因 3 (g3) 前导序列替换 pelB 前导序列并使用单个 lacZ 启动子序列，用短合成寡核苷酸修饰该载体。通过这种方式，L链和Fd的N端氨基酸保持不变，并且基于该新设计的载体构建的随机HCDR3文库没有表现出重组缺失。
Phage display has been used extensively in antibody (Ab) engineering. Sometimes, however, phage display vectors exhibit deletion of immunoglobulin (Ig) genes. As an approach to circumvent the recombination-deletion of the murine anti-digoxin Fab 40–50 cloned into the pComb3 vector, the vector was modified with short synthetic oligonucleotides by replacing a pelB leader sequence with a gene 3 (g3) leader sequence and by using a single lacZ promoter sequence. By this means, the N-terminal amino acids of the L chain and Fd remained unchanged, and a random HCDR3 library built on this newly designed vector did not exhibit the recombination-deletion.