Coupling of antibodies via protein Z on modified polyoma virus-like particles.
Therapeutic application of virus-based delivery systems often implies a change of the tropism of these vectors. This can be achieved by insertion of polypeptides (e.g., antibody fragments) in viral coat proteins. Such fusion proteins have only been used in viral vectors so far and, as part of a virus, they have not been available for a detailed biophysical characterization. We analyzed a fusion protein called VP1-Z, which is based on the polyoma virus coat protein VP1 and protein Z. Protein Z is an engineered antibody-binding domain derived from protein A from Staphylococcus aureus. The fusion VP1-Z was constructed by insertion of protein Z in the HI-loop of VP1. As wild-type VP1, VP1-Z formed pentameric capsomers and assembled to VLPs in vitro. The stability of these particles was very similar compared to that of VLPs of wild-type VP1. Protein Z was fully structured in the fusion protein and was still capable of binding antibodies on the surface of VLPs of VP1-Z. Using this fusion protein, we could change the tropism of polyoma VLPs toward cells presenting on their surface the antigen of the coupled antibody.
Martin-Luther-Universität Halle, Institut für Biotechnologie, D-06120 Halle, Germany.
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D000906:Antibodies / Q000378:metabolism*; D000911:Antibodies, Monoclonal / Q000378:metabolism; D061067:Antibodies, Monoclonal, Humanized; D002272:Carcinoembryonic Antigen / Q000276:immunology / Q000378:metabolism; D002352:Carrier Proteins / Q000378:metabolism*; D002448:Cell Adhesion; D002942:Circular Dichroism; D004926:Escherichia coli / Q000378:metabolism; D050556:Fatty Acid-Binding Proteins; D006388:Hemagglutinins / Q000378:metabolism; D008854:Microscopy, Electron; D008856:Microscopy, Fluorescence; D010957:Plasmids / Q000378:metabolism; D011120:Polyomavirus / Q000276:immunology*; D017433:Protein Structure, Secondary; D017434:Protein Structure, Tertiary; D011993:Recombinant Fusion Proteins / Q000737:chemistry*; D011994:Recombinant Proteins / Q000737:chemistry; D013329:Structure-Activity Relationship; D013696:Temperature; D000068878:Trastuzumab; D014461:Ultracentrifugation
D000906:Antibodies; D000911:Antibodies, Monoclonal; D061067:Antibodies, Monoclonal, Humanized; D002272:Carcinoembryonic Antigen; D002352:Carrier Proteins; D050556:Fatty Acid-Binding Proteins; D006388:Hemagglutinins; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; D000068878:Trastuzumab