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- W2316834962 abstract "Antibody surface display technology is critical for novel antibody screening and antibody affinity maturation. Currently most frequently used display methods are phage display and yeast display. Although Escherichia coli (E. coli) is easily cultured and genetically manipulated, thus is potentially an excellent display host, the display technology based on E. coli has not been widely used. One of the problems is lack of efficient display of antibodies on the surface of E. coli. Many proteins have been tested as display carriers in outer membrane display in E. coli. Display systems based on autotransporter protein (AT) and ice nucleation protein (INP) is the most extensively studied. Another problem is unstable survival rates of E. coli when antibody is displayed. In this study, we systematically examined display level, antigen-binding affinity of displayed antibody and survival rate of E. coli using Ag43茁 (茁 domain of Antigen43, an AT protein) and INPNC (fragment of N-terminal and C-terminal of INP) as carrier proteins and T7, lac, araBAD as promoters for antibody expression. We found that the antigen-binding ability of the Ag43茁 based display was superior to that of the INPNC based system. As expected, T7, lac and araBAD promoters drove high, medium and low expressions of antibody. The host survival rate using T7 promoter was extremely low (INPNC: 0.0033% , Ag43茁: 0.02% , the host bearing araBAD promoter had the highest survival rate (INPNC: 37.80%, Ag43茁: 90.23%), and the lac based system had a survival rate of 2.04% (INPNC) and 13.27% (Ag43茁). Balancing the antigen-binding abilities, antibody expression levels and survival rates, a system using Ag43茁 as carrier protein and lac as promoter is the best choice for antibody display on E. coli." @default.
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- W2316834962 date "2013-01-01" @default.
- W2316834962 modified "2023-10-16" @default.
- W2316834962 title "Comparison of Autotransporter and Ice Nucleation Protein as Carrier Proteins for Antibody Display on The Cell Surface of Escherichia coli" @default.
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- W2316834962 doi "https://doi.org/10.3724/sp.j.1206.2013.00091" @default.
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