Ohmuro-Matsuyama Yuki, Yamaji Hideki
Department of Chemical Science and Engineering, Graduate School of Engineering, Kobe University, 1-1 Rokkodai, Nada, Kobe, 657-8501, Japan.
Laboratory for Chemistry and Life Science, Institute for Innovative Research, Tokyo Institute of Technology, Yokohama, 226-8503, Japan.
Cytotechnology. 2018 Jun;70(3):891-898. doi: 10.1007/s10616-017-0109-0. Epub 2017 Jun 5.
Monoclonal antibodies and antibody fragments have recently been developed for use in diverse diagnostic and therapeutic applications. Insect cells can efficiently secrete recombinant proteins such as antibody molecules through post-translational processing and modifications that are similar to those performed in mammalian cells. In eukaryotic cells, the signal sequence in a nascent polypeptide is recognized by the signal recognition particle, and the polypeptide is then folded and modified in the endoplasmic reticulum. The signal sequence consists of three regions, a positively charged N-terminus, a hydrophobic core, and a polar C-terminus. In the present study, we examined the substitutions of the characteristic amino acids of a Drosophila immunoglobulin heavy chain binding protein signal sequence, and investigated the effect on the secretory production of an antibody Fab fragment from lepidopteran insect cells in transient expression. A modification of the signal sequence for the heavy chain resulted in a twofold increase in the secreted Fab fragment, while the modification for the light chain led to a more than 3.6-fold increase.
单克隆抗体和抗体片段最近已被开发用于各种诊断和治疗应用。昆虫细胞可以通过与哺乳动物细胞中进行的翻译后加工和修饰相似的方式，高效分泌重组蛋白，如抗体分子。在真核细胞中，新生多肽中的信号序列被信号识别颗粒识别，然后多肽在内质网中折叠和修饰。信号序列由三个区域组成，带正电荷的N端、疏水核心和极性C端。在本研究中，我们研究了果蝇免疫球蛋白重链结合蛋白信号序列特征氨基酸的替换，并研究了其对鳞翅目昆虫细胞瞬时表达中抗体Fab片段分泌产生的影响。重链信号序列的修饰使分泌的Fab片段增加了两倍，而轻链的修饰则使增加超过3.6倍。
