Bacterial Display Technology

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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 © 2018 Creative-Biolabs All Rights Reserved Email: [email protected] Tel: 1-631-381-2994.

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Peptide Tag Systems That Spontaneously Form An
(19) TZZ ¥_T (11) EP 2 534 484 B1 (12) EUROPEAN PATENT SPECIFICATION (45) Date of publication and mention (51) Int Cl.: of the grant of the patent: G01N 33/531 (2006.01) C07K 14/315 (2006.01) 19.11.2014 Bulletin 2014/47 C12N 15/00 (2006.01) (21) Application number: 11706621.7 (86) International application number: PCT/GB2011/000188 (22) Date of filing: 11.02.2011 (87) International publication number: WO 2011/098772 (18.08.2011 Gazette 2011/33) (54) PEPTIDE TAG SYSTEMS THAT SPONTANEOUSLY FORM AN IRREVERSIBLE LINK TO PROTEIN PARTNERS VIA ISOPEPTIDE BONDS PEPTIDMARKIERUNGSSYSTEME MIT SPONTANER BILDUNG EINER IRREVERSIBLEN VERBINDUNG ZU PROTEINPARTNERN ÜBER ISOPEPTIDBINDUNGEN SYSTÈMES DE MARQUAGE PEPTIDIQUE QUI FORMENT SPONTANÉMENT UNE LIAISON IRRÉVERSIBLE AVEC DES PARTENAIRES PROTÉIQUES PAR L’INTERMÉDIAIRE DE LIAISONS ISOPEPTIDIQUES (84) Designated Contracting States: • TOMINAGA J ET AL: "Design of a specific peptide AL AT BE BG CH CY CZ DE DK EE ES FI FR GB tagthat affords covalent and site- specificenzyme GR HR HU IE IS IT LI LT LU LV MC MK MT NL NO immobilization catalyzed by microbial PL PT RO RS SE SI SK SM TR transglutaminase", BIOMACROMOLECULES JULY/AUGUST 2005 AMERICAN CHEMICAL (30) Priority: 11.02.2010 GB 201002362 SOCIETY US, vol. 6, no. 4, July 2005 (2005-07), pages 2299-2304, XP002631438, DOI: DOI: (43) Date of publication of application: 10.1021/BM050193O 19.12.2012 Bulletin 2012/51 • DONG RUI-PING ET AL: "Characterization of T cell epitopes restricted by HLA-DP9 in (73) Proprietor: Isis Innovation Limited streptococcal M12 protein", JOURNAL OF Summertown, Oxford OX2 7SQ (GB) IMMUNOLOGY, vol.
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