Staphylococcal surface display of immunoglobulin A (IgA)- and IgE-specific in vitro-selected binding proteins (affibodies) based on Staphylococcus aureus protein A.
An expression system designed for cell surface display of hybrid proteins on Staphylococcus carnosus has been evaluated for the display of Staphylococcus aureus protein A (SpA) domains, normally binding to immunoglobulin G (IgG) Fc but here engineered by combinatorial protein chemistry to yield SpA domains, denoted affibodies, with new binding specificities. Such affibodies, with human IgA or IgE binding activity, have previously been selected from a phage library, based on an SpA domain. In this study, these affibodies have been genetically introduced in monomeric or dimeric forms into chimeric proteins expressed on the surface of S. carnosus by using translocation signals from a Staphylococcus hyicus lipase construct together with surface-anchoring regions of SpA. The recombinant surface proteins, containing the IgA- or IgE-specific affibodies, were demonstrated to be expressed as full-length proteins, localized and properly exposed at the cell surface of S. carnosus. Furthermore, these chimeric receptors were found to be functional, since recombinant S. carnosus cells were shown to have gained IgA and IgE binding capacity, respectively. In addition, a positive effect in terms of IgA and IgE reactivity was observed when dimeric versions of the affibodies were present. Potential applications for recombinant bacteria with redirected binding specificity in their surface proteins are discussed.
Gunneriusson
E
E
Department of Biotechnology, Royal Institute of Technology (KTH), S-100 44 Stockholm, Sweden.
Samuelson
P
P
Ringdahl
J
J
Grönlund
H
H
Nygren
P A
PA
Ståhl
S
S
D000907:Antibodies, Bacterial / Q000096:biosynthesis / Q000276:immunology; D000954:Antigens, Surface / Q000276:immunology*; D006801:Humans; D007070:Immunoglobulin A / Q000276:immunology*; D007073:Immunoglobulin E / Q000276:immunology*; D011993:Recombinant Fusion Proteins / Q000737:chemistry / Q000276:immunology*; D013205:Staphylococcal Protein A / Q000737:chemistry / Q000276:immunology*; D013210:Staphylococcus / Q000737:chemistry / Q000235:genetics / Q000276:immunology*
D000907:Antibodies, Bacterial; D000954:Antigens, Surface; D007070:Immunoglobulin A; D011993:Recombinant Fusion Proteins; D013205:Staphylococcal Protein A; D007073:Immunoglobulin E