Integral membrane protein display on poxvirus extracellular enveloped virions
AssigneeVaccinex, Inc.
InventorsErnest S. Smith, Mark Paris, Maria G. M. Scrivens, Renee A. Kirk, Angelica A. Cornelison
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Abstract
This disclosure provides compositions and methods for expressing and displaying isolated integral membrane proteins (IMPs) or fragments thereof in a native conformation for use in the screening, selecting, and identifying of antibodies or antibody-like molecules that bind to a target IMP of interest.
Claims15
An isolated polynucleotide comprising:
- (a) a first nucleic acid fragment that encodes an integral membrane protein (IMP) or fragment thereof, wherein the IMP or fragment thereof comprises at least one extra-membrane region, at least one transmembrane domain and at least one intra-membrane region, and wherein a portion of the first nucleic acid fragment encoding at least one intra-membrane region is situated at the 5′ or 3′ end of the first nucleic acid fragment;
- and
- (b) a second nucleic acid fragment that encodes a poxvirus EEV-specific protein or functional fragment thereof, wherein the second nucleic acid fragment is fused in frame to a portion of the first nucleic acid fragment that encodes an intra-membrane region of the IMP;
- wherein a poxvirus infected cell comprising the polynucleotide can express an IMP-poxvirus EEV-specific protein fusion protein as part of the outer envelope membrane of an extracellular enveloped virion (EEV).
A recombinant poxvirus EEV comprising a heterologous IMP or fragment thereof fused to a poxvirus EEV-specific protein or membrane-associated fragment thereof, wherein the fusion protein is situated in the EEV outer envelope membrane, wherein the IMP or fragment thereof displays on the surface of the EEV in its native conformation.
Dependencies are inferred from the claim text.