Bacterial Display Technology

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 bacteria 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 bacteria 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