CD40-Ligand lacking native-pattern glycosylation
AssigneeImmunex Corporation
InventorsRichard J. Armitage, William C. Fanslow, III, Melanie K. Spriggs
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Abstract
There is disclosed a polypeptide (CD40-L) and DNA sequences, vectors and transformed host cells useful in providing CD40-L polypeptides. More particularly, this invention provides isolated human and murine CD40-L polypeptides that bind to the extracellular binding region of a CD40 receptor and lack native-pattern glycosylation.
Claims27
A soluble C40-L polypeptide (CD40-L) that binds CD40 and lacks a CD40-L tranamembrane region, wherein the CD40-L polypeptide is encoded by a nucleic acid selected from the group consisting of:
- (a) a polynucleotide comprising nucleotides 184 through 828 of SEQ ID NO:11;
- (b) a polynucleotide comprising nucleotides 196 through 828 of SEQ ID NO:11;
- (c) a polynucleotide comprising nucleotides 403 through 828 of SEQ ID NO:11;
- (d) a polynucleotide comprising nucleotides 382 through 828 of SEQ ID NO:11;
- and
- (e) a polynucleotide comprising nucleotides 379 through 828 of SEQ ID NO:11;
- wherein the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, wherein the CD40-L polypeptide is encoded by a nucleic acid selected from the group consisting of:
- (a) a polynucleotide comprising nucleotides 46 through 828 of SEQ ID NO:11;
- (b) a polynucleotide comprising nucleotides 184 through 828 of SEQ ID NO:11;
- (c) a polynucleotide comprising nucleotides 196 through 828 of SEQ ID NO:11;
- (d) a polynucleotide comprising nucleotides 403 through 828 of SEQ ID NO:11;
- (e) a polynucleotide comprising nucleotides 382 through 828 of SEQ ID NO:11;
- (f) a polynucleotide comprising nucleotides 379 through 828 of SEQ ID NO:11;
- and
- (g) a polynucleotide that hybridizes to the complement of a polynucleotide of (a)-(f) under moderately stringent conditions (prewashing solution of 5×SSC, 05% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight),
- wherein the nucleotides 625 to 627 of SEQ ID NO:11 are substituted to encode tryptophan instead of cysteine and the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40selected from the group consisting of:
- (a) a polypeptide comprising amino acids 1 through 261 of SEQ ID NO:12;
- (b) a polypeptide comprising amino acids 47 through 261 of SEQ ID NO:12;
- (c) a polypeptide comprising amino acids 51 through 261 of SEQ ID NO:12;
- (d) a polypeptide comprising amino acids 120 through 261 of SEQ ID NO:12;
- (e) a polypeptide comprising amino acids 113 through 261 of SEQ ID NO:12;
- and
- (f) a polypeptide comprising amino acids 112 through 261 of SEQ ID NO:12,
- wherein the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide selected from the group consisting of
- (a) a polypeptide comprising amino acids 1 through 261 of SEQ ID NO:12;
- (b) a polypeptide comprising amino acids 47 through 261 of SEQ ID NO:
- 12;
- (c) a polypeptide comprising amino acids 51 through 261 of SEQ ID NO:12;
- (d) a polypeptide comprising amino acids 120 through 261 of SEQ ID NO:12;
- (e) a polypeptide comprising amino acids 113 through 261 of SEQ ID NO:12;
- and
- (f) a polypeptide comprising amino acids 112 through 261 of SEQ ID NO:12,
- wherein the cysteine at amino acid 194 of SEQ ID NO:12 is substituted with tryptophan and the CD40-L polypeptide lacks native-pattern glycosylation.
Dependencies are inferred from the claim text.