title: An autotransporter display platform for the development of multivalent recombinant bacterial vector vaccines.
abstract: BACKGROUND
  The Autotransporter pathway, ubiquitous in Gram-negative bacteria, allows the efficient secretion of large passenger proteins via a relatively simple mechanism. Capitalizing on its crystal structure, we have engineered the Escherichia coli autotransporter Hemoglobin protease (Hbp) into a versatile platform for secretion and surface display of multiple heterologous proteins in one carrier molecule.
  
  
  RESULTS
  As proof-of-concept, we demonstrate efficient secretion and high-density display of the sizeable Mycobacterium tuberculosis antigens ESAT6, Ag85B and Rv2660c in E. coli simultaneously. Furthermore, we show stable multivalent display of these antigens in an attenuated Salmonella Typhimurium strain upon chromosomal integration. To emphasize the versatility of the Hbp platform, we also demonstrate efficient expression of multiple sizeable antigenic fragments from Chlamydia trachomatis and the influenza A virus at the Salmonella cell surface.
  
  
  CONCLUSIONS
  The successful efficient cell surface display of multiple antigens from various pathogenic organisms highlights the potential of Hbp as a universal platform for the development of multivalent recombinant bacterial vector vaccines.
authors.affiliation: Department of Molecular Cell Biology, Section Molecular Microbiology, Faculty of Earth and Life Sciences, VU University, De Boelelaan 1085, 1081, HV, Amsterdam, The Netherlands. w.s.p.jong@vu.nl.|Abera Bioscience AB, SE-111 45, Stockholm, Sweden. w.s.p.jong@vu.nl.
authors.forename: Wouter S P
authors.initials: WS
authors.lastname: Jong
authors.affiliation: Department of Molecular Cell Biology, Section Molecular Microbiology, Faculty of Earth and Life Sciences, VU University, De Boelelaan 1085, 1081, HV, Amsterdam, The Netherlands. maria.daleke@aberabio.com.|Abera Bioscience AB, SE-111 45, Stockholm, Sweden. maria.daleke@aberabio.com.
authors.forename: Maria H
authors.initials: MH
authors.lastname: Daleke-Schermerhorn
authors.affiliation: Xbrane Bioscience AB, SE-111 45, Stockholm, Sweden. david.vikstrom@xbrane.com.|Department of Biochemistry and Biophysics, Center for Biomembrane Research, Stockholm University, SE-106 91, Stockholm, Sweden. david.vikstrom@xbrane.com.
authors.forename: David
authors.initials: D
authors.lastname: Vikström
authors.affiliation: Department of Molecular Cell Biology, Section Molecular Microbiology, Faculty of Earth and Life Sciences, VU University, De Boelelaan 1085, 1081, HV, Amsterdam, The Netherlands. c.m.ten.hagen-jongman@vu.nl.|Abera Bioscience AB, SE-111 45, Stockholm, Sweden. c.m.ten.hagen-jongman@vu.nl.
authors.forename: Corinne M
authors.initials: CM
authors.lastname: Ten Hagen-Jongman
authors.affiliation: The Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital, 1066, CX, Amsterdam, The Netherlands. karin.de-punder@charite.de.|Present Address: Institute for Medical Psychology, Charité Universitätsmedizin, 10117, Berlin, Germany. karin.de-punder@charite.de.
authors.forename: Karin
authors.initials: K
authors.lastname: de Punder
authors.affiliation: The Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital, 1066, CX, Amsterdam, The Netherlands. N.N.vanderWel@amc.uva.nl.|Present Address: Department of Cell Biology and Histology, Academic Medical Center, University of Amsterdam, 1105, AZ, Amsterdam, The Netherlands. N.N.vanderWel@amc.uva.nl.
authors.forename: Nicole N
authors.initials: NN
authors.lastname: van der Wel
authors.affiliation: Department of Viroscience, Erasmus Medical Center, 3015, GE, Rotterdam, The Netherlands. c.vandesandt@erasmusmc.nl.
authors.forename: Carolien E
authors.initials: CE
authors.lastname: van de Sandt
authors.affiliation: Department of Viroscience, Erasmus Medical Center, 3015, GE, Rotterdam, The Netherlands. g.rimmelzwaan@erasmusmc.nl.
authors.forename: Guus F
authors.initials: GF
authors.lastname: Rimmelzwaan
authors.affiliation: Department of Infectious Disease & Immunology, Statens Serum Institut, Copenhagen, Denmark. FRF@ssi.dk.
authors.forename: Frank
authors.initials: F
authors.lastname: Follmann
authors.affiliation: Department of Infectious Disease & Immunology, Statens Serum Institut, Copenhagen, Denmark. EAG@ssi.dk.
authors.forename: Else Marie
authors.initials: EM
authors.lastname: Agger
authors.affiliation: Department of Infectious Disease & Immunology, Statens Serum Institut, Copenhagen, Denmark. PA@ssi.dk.
authors.forename: Peter
authors.initials: P
authors.lastname: Andersen
authors.affiliation: Xbrane Bioscience AB, SE-111 45, Stockholm, Sweden. degier@dbb.su.se.|Department of Biochemistry and Biophysics, Center for Biomembrane Research, Stockholm University, SE-106 91, Stockholm, Sweden. degier@dbb.su.se.
authors.forename: Jan-Willem
authors.initials: JW
authors.lastname: de Gier
authors.affiliation: Department of Molecular Cell Biology, Section Molecular Microbiology, Faculty of Earth and Life Sciences, VU University, De Boelelaan 1085, 1081, HV, Amsterdam, The Netherlands. s.luirink@vu.nl.|Abera Bioscience AB, SE-111 45, Stockholm, Sweden. s.luirink@vu.nl.
authors.forename: Joen
authors.initials: J
authors.lastname: Luirink
mesh_terms: D000942:Antigens, Bacterial / Q000235:genetics / Q000378:metabolism; D000956:Antigens, Viral / Q000235:genetics / Q000378:metabolism; D058947:Bacterial Secretion Systems; D001428:Bacterial Vaccines / Q000235:genetics / Q000378:metabolism; D002692:Chlamydia trachomatis / Q000235:genetics; D010450:Endopeptidases / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics / Q000378:metabolism; D009980:Influenza A virus / Q000235:genetics; D007252:Influenza Vaccines / Q000235:genetics / Q000378:metabolism; D009169:Mycobacterium tuberculosis / Q000235:genetics; D012486:Salmonella typhimurium / Q000235:genetics / Q000378:metabolism
chemical_list: D000942:Antigens, Bacterial; D000956:Antigens, Viral; D058947:Bacterial Secretion Systems; D001428:Bacterial Vaccines; D007252:Influenza Vaccines; D010450:Endopeptidases; C114676:hemoglobin protease Hbp