Method for exposing peptides and polypeptides on the cell...

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

Reexamination Certificate

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C435S252300, C435S320100

Reexamination Certificate

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10415165

ABSTRACT:
The inventive method allows peptides or polypeptides to be exposed on the surface of gram-negative host bacteria using specific intimin-based anchor modules. Intimins with shortened carboxy terminals have been found to be particularly suitable anchor modules for passenger domains in the exteriorE. colicell membrane. According to the method, host bacteria are transformed using vectors, on which are located a fused nucleic acid sequence consisting of a sequence segment which codes for an intimin with a shortened carboxy terminal and a nucleic acid sequence segment which codes for the passenger peptide that is to be exposed. The invention permits a particularly large number of passenger domains to be exposed on the cell surface of the bacteria, without adversely affecting the viability of the bacteria.

REFERENCES:
patent: 0 872 555 (1997-02-01), None
patent: WO 97/40177 (1997-10-01), None
Candy et al.; Characterization of the C-terminal domains of intimin-like proteins of enteropathogenic and enterohemorrhagicEscherichia coli, Citrobacter freundii, andHafnia alvei; dated May 1994 Infection + Immunity 62 (5) pp. 1835-1842.
Christmann et al. The cystine knot of a squash -type protease inhibitor as a structural scaffold forEscherichia colicell surface display of conformationally constrained peptides.Protein Engineering vol. 12. No. 9 pp. 797-806; dated 1999.
Schembri et al. XP-002109166′; Applied and Enviromental Microbiology, May 1998 pp. 1628-1633; vol. 64 No. 5 , Heterobinary Adhesins Based on theEscherichia coliFim H Fimbrial Protein.
DNA Circular; XP-002211330; Expression vector for surface display of Intimin′ fusion proteins on cells ofEscherichia coli; dated Sep. 11, 2001.
Christmann et al. Journal of Immunological Methods 257 (2001) 163-173 Epitope mapping and affinity purification of monospecific antibodies byEscherichia colicell surface display of gene derived random peptide libraries. dated Jul. 8, 2001.
Journal of Bacteriology, Dec. 2001 pp. 7273-7284 Wenzel et al., XP-002211329; Display of passenger proteins on the surface ofEscherichia coliK-12 by the EnterohemorrhagicE. coliIntimin EaeA.
Nature Structural Biology, vol. 6, No. 6; Kelly et al. XP-001018419Structure of the cell- adhesion fragment of intimin from enteropathogenicEscherichia coli; dated Apr. 1999 pp. 313-318.

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