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  • 03월 02일 17시 이후 : 초록수정 불가능, 일정확인 및 검색만 가능

제109회 대한화학회 학술발표회, 총회 및 기기전시회 안내 Tailored preparation of E.coli outer membrane with autodisplayed proteins for immunoassays

2012년 2월 16일 15시 38분 24초
BIO.P-683 이곳을 클릭하시면 발표코드에 대한 설명을 보실 수 있습니다.
4월 25일 (수요일) 18:00~21:00
저자 및
박민, 변재철1
연세대학교 신소재공학과, Korea
1연세대학교 신소재공학부, Korea
The autodisplay technology is one of the surface display methods for proteins or peptides at the E.coli outer membrane. In the autodisplay technology, proteins or peptides are expressed as recombinant proteins in a poly protein precursor for the autotransporter secretion pathway and automatically aligned on the outer membrane surface. These aligned proteins or peptides can be used as a molecular recognition layer or a affinity layer for immunoassay. For the preparation of outer membrane from autodiaplyed E.coli, lysozyme was treated to hydrolyze the peptidoglycan layer and triton X-100 was used for dissolving the cell membrane. After the outer membrane isolation, marker proteins from outer membrane, periplasm, inner membrane and cytoplasm were selected and analyezed by western blot and the enzymatic assay. OmpF, β-lactamase, secA and β-galactosidase were used for western blot and KDO, β-lactamase, NADH oxidase and β-galactosidase were analyzed by enzymatic assay. Isolated outer membrane were layered on the surface of 96 well microplate by using hydrophobic interaction and layered outer membrane were analyezed by enzymatic assay. The purity of isolated outer membrane and its layer was calculated and the Z-domains, have a binding affinity to Fc region of IgG, autodisplaying outer membrane layer was used for the immunoassay to improve the sensitivity.