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US 10,550,199 B2Granted
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
Life of this patent
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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.
Claims24
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 vaccinia virus F13L protein comprising the amino acid sequence SEQ ID NO:
- 1 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-F13L fusion protein as part of the outer envelope membrane of an extracellular enveloped virion (EEV).
2 more elements
A method to display an integral membrane protein (IMP) or fragment thereof in a native conformation comprising:
- (a) infecting host cells permissive for poxvirus infectivity with a recombinant poxvirus that expresses the IMP or fragment thereof as a fusion protein with poxvirus EEV-specific protein or membrane-associated fragment thereof, wherein EEV produced by the infected host cell comprise the IMP fusion protein as part of the EEV outer envelope membrane;
- (b) recovering EEV released from the host cell;
- wherein the IMP or fragment thereof displays on the surface of the EEV in a native conformation.
Dependencies are inferred from the claim text.