Variant forms of urate oxidase and use thereof
First Claim
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1. A method for producing a uricase comprising the steps of:
- (i) culturing a host cell comprising a vector comprising a nucleic acid sequence encoding a uricasecomprising a mammalian uricase amino acid sequence truncated at the amino terminus or the carboxy terminus or both the amino and carboxy termini by 1-6 amino acids and further comprising an amino acid substitution with threonine at position 7 (D7T), an amino acid substitution at position 46 (S46T), an amino acid substitution with lysine at position 291 (R291K) and an amino acid substitution with serine at position 301 (T301S), said truncation and amino acid substitution being relative to an amino acid sequence of SEQ ID NO;
11;
wherein the nucleic acid sequence is operatively linked to a heterologous promoter under conditions such that the nucleic acid sequence is expressed by the host cell; and
(ii) isolating the expressed uricase.
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Abstract
Genetically modified proteins with uricolytic activity are described. Proteins comprising truncated urate oxidases and methods for producing them, including PEGylated proteins comprising truncated urate oxidase are described.
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Citations
17 Claims
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1. A method for producing a uricase comprising the steps of:
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(i) culturing a host cell comprising a vector comprising a nucleic acid sequence encoding a uricase comprising a mammalian uricase amino acid sequence truncated at the amino terminus or the carboxy terminus or both the amino and carboxy termini by 1-6 amino acids and further comprising an amino acid substitution with threonine at position 7 (D7T), an amino acid substitution at position 46 (S46T), an amino acid substitution with lysine at position 291 (R291K) and an amino acid substitution with serine at position 301 (T301S), said truncation and amino acid substitution being relative to an amino acid sequence of SEQ ID NO;
11;wherein the nucleic acid sequence is operatively linked to a heterologous promoter under conditions such that the nucleic acid sequence is expressed by the host cell; and (ii) isolating the expressed uricase. - View Dependent Claims (2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17)
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Specification