Non-hemolytic LLO fusion proteins and methods of utilizing same
First Claim
1. A recombinant protein comprising a listeriolysin O (LLO) protein or N-terminal fragment thereof, wherein said LLO protein or N-terminal fragment comprises a mutation in a cholesterol-binding domain (CBD), wherein said mutation comprises:
- a. a substitution of 1-50 μ
amino acid peptide in said CBD as set forth in SEQ ID NO;
18 for a 1-50 amino acid non-LLO peptide;
b. a mutation of residue C484, W491, or W492 of SEQ ID NO;
37 or a combination thereof;
orc. a deletion of a 1-50 amino acid peptide in said CBD as set for in SEQ ID NO;
18.wherein said recombinant protein exhibits a greater than 100-fold reduction in hemolytic activity relative to wild-type LLO.
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Accused Products
Abstract
The present invention provides recombinant proteins or peptides comprising a mutated listeriolysin O (LLO) protein or fragment thereof, comprising a substitution or internal deletion of the cholesterol-binding domain or a portion thereof, fusion proteins or peptides comprising same, nucleotide molecules encoding same, and vaccine vectors comprising or encoding same. The present invention also provides methods of utilizing recombinant proteins, peptides, nucleotide molecules, and vaccine vectors of the present invention to induce an immune response to a peptide of interest.
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Citations
61 Claims
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1. A recombinant protein comprising a listeriolysin O (LLO) protein or N-terminal fragment thereof, wherein said LLO protein or N-terminal fragment comprises a mutation in a cholesterol-binding domain (CBD), wherein said mutation comprises:
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a. a substitution of 1-50 μ
amino acid peptide in said CBD as set forth in SEQ ID NO;
18 for a 1-50 amino acid non-LLO peptide;b. a mutation of residue C484, W491, or W492 of SEQ ID NO;
37 or a combination thereof;
orc. a deletion of a 1-50 amino acid peptide in said CBD as set for in SEQ ID NO;
18.wherein said recombinant protein exhibits a greater than 100-fold reduction in hemolytic activity relative to wild-type LLO. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, 60)
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30-59. -59. (canceled)
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61-81. -81. (canceled)
Specification