제목 | [논문] 이상호 교수님_Antibody reformatting vector system (AbVec): a platform for converting single-chain variable fragment... | ||||
---|---|---|---|---|---|
작성자 | 비임파성 장기 면역 연구센터 | 등록일 | 2021-01-22 | 조회수 | 885 |
Antibody reformatting vector system (AbVec): a platform for converting single-chain variable fragment format to other ones using shared restriction enzyme sites
Single-chain antibody variable fragment (scFv) is widely used at screening stage. However, the scFv format often proves tobe inefficient in downstream stages such as crystallization and in vivo applications. Conversion of a scFv format to otherones such as a fragment antigen binding (Fab), a scFv-fragment crystallizable region (scFv-Fc), a scFv-constant heavychain (scFv-CH) and an immunoglobulin (IgG) formats can be cumbersome due to different sets of restriction enzyme sitesin multiple vectors. To facilitate antibody format conversion in laboratory settings, we developed a cloning platform, called“AbVec”, with the common restriction enzyme sites across various forms of vectors. In the AbVec platform, a scFv cloneselected from phage display is modularized into VH and VL regions and transferred to vectors for Fab, scFv-Fc, scFv-CH and IgG production using the same restriction enzyme sites. Using scFv F9, specifically recognizing a pneumococcalsecreted peptide, we demonstrated the AbVec platform yielded the expression vectors within a week. Different formats ofthe antibodies were successfully expressed and purified homogenously, supporting the usefulness of the AbVec platform. The AbVec platform will facilitate otherwise time-consuming conversion process in laboratory settings. |