The 129th Annual Meeting of Japanese Society of Animal Science

Presentation information

口頭発表

5. Animal products technology

畜産物利用2

Wed. Sep 15, 2021 1:00 PM - 2:30 PM 畜産物利用 (オンライン)

Chairperson:Shiro Takeda, Suguru Shigemori, Binghui Zhou

[V-15-25] Construction of genetically modified lactic acid bacteria producing anti-IL31Ra single-chain Fragment variable

*Aito Murakami1, Fu Namai1,2, Suguru Shigemori1, Tasuku Ogita1, Takeshi Shimosato1 (1. IBS., Shinshu Univ., 2. Graduate School of Biomedical Sciences, Nagasaki Univ.)

[Objectives] In order to suggest the further application of lactic acid bacteria (LAB), we conducted research on genetically modified LAB (gmLAB). Here, we constructed a gmLAB producing anti-IL31 receptor A single-chain fragment variable (IL31RascFv) for atopic dermatitis treatment and evaluated the binding ability of the scFv. [Methods] The gene expression vector with IL31RascFv sequences was introduced into Lactococcus lactis NZ9000 to generate a gmLAB (designated as NZ-IL31RascFv). The expression analysis of IL31RascFv was performed using Western Blot (WB) and the immunoreactivity of scFv against antigen protein, IL-31 Ra, was investigated using ELISA. [Results] We confirmed a band corresponding to the IL31RascFv by WB. During ELISA, the absorbance increased in a concentration-dependent manner when NZ-IL31RascFv cellular extract was added to the well immobilized with IL-31 Ra. Therefore, we successfully constructed a gmLAB-expressing immunoreactive IL31RascFv.