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.
Claims26
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40L polypeptide, wherein said nucleic acid hybridizes to a polynucleotide selected from the group consisting of (a)-(f), or its complement, under moderately stringent conditions (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (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;
- and
- (f) a polynucleotide comprising nucleotides 379 through 828 of SEQ ID NO:11,
- with the proviso that the CD40-L lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40L polypeptide, wherein said nucleic acid hybridizes to a polynucleotide selected from the group consisting of (a)-(f), or its complement, under moderately stringent conditions (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (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 polynucleotidecomprising nucleotides 403 through 828 of SEQ ID NO:11;
- (e) a polynucleotide comprising nucleotides 382 through 828 of SEQ ID NO:11;
- and
- (f) a polynucleotide comprising nucleotides 379 through 828 of SEQ ID NO:11,
- with the provisos that the nucleotides encoding cysteine at nucleotides 625 to 627 of SEQ ID NO:
- 11 are substituted with DNA encoding tryptophan and that the CD40-L lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid that encodes the CD40-L polypeptide, comprising nucleotides 379 through 828 of SEQ ID NO:11 wherein the DNA encoding cysteine at nucleotides 625 to 627 of SEQ ID NO:11 is substituted with DNA encoding tryptophan, and further comprising a DNA molecule encoding a leucine zipper as set forth in SEQ ID NO:17, with the proviso that the CD40-L lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40-L polypeptide, wherein the nucleic acid hybridizes to the complement of a nucleic acid that encodes a CD40L polypeptide selected from the group consisting of polypeptides (a)-(f), and wherein the hybridization conditions are moderately stringent (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (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, with the proviso that the CD40-L lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40-L polypeptide, wherein the nucleic acid hybridizes to the complement of a nucleic acid that encodes a CD40L polypeptide selected from the group consisting of polypeptides (a)-(b), and wherein the hybridization conditions are moderately stringent (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (a) a polypeptide comprising amino acids 1 through 261 of SEQ ID NO:12;
- and
- (b) a polypeptide comprising amino acids 1 through 261 of SEQ ID NO:12, with the provisos that the cysteine at amino acid 194 is substituted with tyrptophan and that the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40-L polypeptide, wherein the nucleic acid hybridizes to the complement of a nucleic acid that encodes a CD40L polypeptide selected from the group consisting of polypeptides (a)-(b), and wherein the hybridization conditions are moderately stringent (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (a) a polypeptide comprising amino acids 47 through 261 of SEQ ID NO:12;
- and
- (b) a polypeptide comprising amino acids 47 through 261 of SEQ ID NO:12, with the provisos that the cysteine at amino acid 194 is substituted with tyrptophan and that the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40-L polypeptide, wherein the nucleic acid hybridizes to the complement of a nucleic acid that encodes a CD40L polypeptide selected from the group consisting of polypeptides (a)-(b), and wherein the hybridization conditions are moderately stringent (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (a) a polypeptide comprising amino acids 51 through 261 of SEQ ID NO:12;
- and
- (b) a polypeptide comprising amino acids 51 through 261 of SEQ ID NO:12, with the provisos that the cysteine at amino acid 194 is substituted with tyrptophan and that the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40-L polypeptide, wherein the nucleic acid hybridizes to the complement of a nucleic acid that encodes a CD40L polypeptide selected from the group consisting of polypeptides (a)-(b), and wherein the hybridization conditions are moderately stringent (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (a) a polypeptide comprising amino acids 120 through 261 of SEQ ID NO:12;
- and
- (b) a polypeptide comprising amino acids 120 through 261 of SEQ ID NO:12, with the provisos that the cysteine at amino acid 194 is substituted with tyrptophan and that the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40-L polypeptide, wherein the nucleic acid hybridizes to the complement of a nucleic acid that encodes a CD40L polypeptide selected from the group consisting of polypeptides (a)-(b), and wherein the hybridization conditions are moderately stringent (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (a) a polypeptide comprising amino acids 113 through 261 of SEQ ID NO:12;
- and
- (b) a polypeptide comprising amino acids 113 through 261 of SEQ ID NO:12, with the provisos that the cysteine at amino acid 194 is substituted with tyrptophan and that the CD40-L polypeptide lacks native-pattern glycosylation.
An isolated CD40-L polypeptide that binds CD40, comprising a nucleic acid encoding the CD40-L polypeptide, wherein the nucleic acid hybridizes to the complement of a nucleic acid that encodes a CD40L polypeptide selected from the group consisting of polypeptides (a)-(b), and wherein the hybridization conditions are moderately stringent (prewashing solution of 5×SSC, 0.5% SDS, 1.0 mM EDTA, (pH 8.0) and hybridization conditions of 50° C., 5×SSC, overnight):
- (a) a polypeptide comprising amino acids 112 through 261 of SEQ ID NO:12;
- and
- (b) a polypeptide comprising amino acids 112 through 261 of SEQ ID NO:12, with the provisos that the cysteine at amino acid 194 is substituted with tyrptophan and that the CD40-L polypeptide lacks native-pattern glycosylation.
Dependencies are inferred from the claim text.