Production of Recombinant Monoclonal Antibodies in the Egg White of Gene-Targeted Transgenic Chickens
-
- Takehiro Mukae
- Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, 1-8-31, Ikeda, Osaka 563-8577, Japan
-
- Sho Okumura
- Sapporo Division, Cosmo Bio Co. Ltd., 3-513-2, Zenibako, Otaru, Hokkaido 047-0261, Japan
-
- Takuma Watanobe
- Sapporo Division, Cosmo Bio Co. Ltd., 3-513-2, Zenibako, Otaru, Hokkaido 047-0261, Japan
-
- Kyoko Yoshii
- Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, 1-8-31, Ikeda, Osaka 563-8577, Japan
-
- Takahiro Tagami
- Animal Breeding and Reproduction Research Division, Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization, 2 Ikenodai, Tsukuba, Ibaraki 305-0901, Japan
-
- Isao Oishi
- Biomedical Research Institute, National Institute of Advanced Industrial Science and Technology, 1-8-31, Ikeda, Osaka 563-8577, Japan
説明
<jats:p>Increased commercial demand for monoclonal antibodies (mAbs) has resulted in the urgent need to establish efficient production systems. We previously developed a transgenic chicken bioreactor system that effectively produced human cytokines in egg whites using genome-edited transgenic chickens. Here, we describe the application of this system to mAb production. The genes encoding the heavy and light chains of humanized anti-HER2 mAb, linked by a 2A peptide sequence, were integrated into the chicken ovalbumin gene locus using a CRISPR/Cas9 protocol. The knock-in hens produced a fully assembled humanized mAb in their eggs. The mAb expression level in the egg white was 1.4–1.9 mg/mL, as determined by ELISA. Furthermore, the antigen binding affinity of the anti-HER2 mAb obtained was estimated to be equal to that of the therapeutic anti-HER2 mAb (trastuzumab). In addition, antigen-specific binding by the egg white mAb was demonstrated by immunofluorescence against HER2-positive and -negative cells. These results indicate that the chicken bioreactor system can efficiently produce mAbs with antigen binding capacity and can serve as an alternative production system for commercial mAbs.</jats:p>
収録刊行物
-
- Genes
-
Genes 12 (1), 38-, 2020-12-30
MDPI AG