BACTERIAL DISPLAY TECHNOLOGY 1 Bacterial display systems Multiple cell surface display platforms on bacteria have - - - been established. For display G bacteria G bacteria G bacteria on Gram-negative bacteria, Insertion N-terminal C-terminal insertion scaffolds (based on scaffolds: N display display C OmpA, OmpX, FhuA, FliTrx, OmpA scaffolds: scaffolds: FimH), N-terminal display OmpX EstA Lpp-OmpA scaffolds (based on EstA, FhuA AIDA-I Invasin N AIDA-I, IgA protease), C-ter- N C FliTrx C IgA protease minal display scaffolds (based FimH on Lpp-OmpA, Invasin), and combination of N-terminal and C-terminal display using - - circularly permuted OmpX G bacteria G bacteria G+ bacteria: (CPX) could be chosen. For N-terminal C-terminal display via Gram-positive bacteria, N C display N C display covalently foreign proteins could be using using N anchored displayed via covalently CPX CPX proteins anchored proteins including C SPA SPA and FnBTB. FnBTB 2 Construction and screening of display library The protein or peptide library is constructed using recombi- nant DNA technology and displayed on bacteria. The bacterial library can be Library DNA labeled with fluorescently Lower-affinity variants Cloning Lower-affinity labeled target protein variants (antigen), or incubated with biotin-tagged target protein and streptavidin-magnetic beads. Target-binding bacte- ria are isolated using fluores- FACS MACS cence-activated cell sorting Highest-affinity Plasmid constructs Highest-affinity (FACS) or magnetic-activated variant variant cell sorting (MACS). The selection cycle is generally repeated, and selected bacte- ria are typically amplified by growth in between the cycles. E. coli Finally, isolated binders are screened using whole-cell Amplification Amplification ELISA-based methods, identi- fied using DNA sequencing and subcloned for soluble production and detailed characterization. Biotin-tagged Fluorescently Display library reporter protein & labeled reporter protein streptavidin-magnetic beads WHAT WE DO: Bacterial Display Services Creative Biolabs Bacterial Display Library Construction Related Services Bacterial Display Epitope Mapping by Bacterial Display Ribosome Display Services Services Cell-Binding Peptides Identification Yeast Display Services Protease Substrate Identification Mammalian Cell Display Service Protein-Binding Ligand Screening Phage Display & Antibody Library Service © 2018 Creative-Biolabs All Rights Reserved Email: [email protected] Tel: 1-631-381-2994.