PURE ribosome display and its application in antibody technology |
| |
Authors: | Takashi Kanamori Yasuhiro Fujino Takuya Ueda |
| |
Affiliation: | 1. The Department of Medical Genome Sciences, Graduate School of Frontier Sciences, The University of Tokyo, FSB-401, 5-1-5, Kashiwanoha, Kashiwa, Chiba 277-8562, Japan;2. Advanced Medical Research Department, Mitsubishi Tanabe Pharma Corporation, 1000 Kamosida, Aoba-ku, Yokohama 227-0033, Japan |
| |
Abstract: | Ribosome display utilizes formation of the mRNA–ribosome–polypeptide ternary complex in a cell-free protein synthesis system to link genotype (mRNA) to phenotype (polypeptide). However, the presence of intrinsic components, such as nucleases in the cell-extract-based cell-free protein synthesis system, reduces the stability of the ternary complex, which would prevent attainment of reliable results. We have developed an efficient and highly controllable ribosome display system using the PURE (Protein synthesis Using Recombinant Elements) system. The mRNA–ribosome–polypeptide ternary complex is highly stable in the PURE system, and the selected mRNA can be easily recovered because activities of nucleases and other inhibitory factors are very low in the PURE system. We have applied the PURE ribosome display to antibody engineering approaches, such as epitope mapping and affinity maturation of antibodies, and obtained results showing that the PURE ribosome display is more efficient than the conventional method. We believe that the PURE ribosome display can contribute to the development of useful antibodies. This article is part of a Special Issue entitled: Recent advances in molecular engineering of antibody. |
| |
Keywords: | CDR, complementarity determining region ELISA, enzyme-linked immunosorbent assay ER, enrichment ratio Fab, fragment antigen binding KinExA, kinetic exclusion assay Kd, dissociation constant ORF, open reading frame PURE system, Protein synthesis Using Recombinant Elements system RD, ribosome display scFv, single-chain Fv SPR, surface plasmon resonance |
本文献已被 ScienceDirect 等数据库收录! |
|