Immunex Corporation

Michael C Deeley

Virginia L Price

David L Urdal

Steven D Gimpel

An analog human colony stimulating factor (hCSF) is disclosed, comprising a mutant amino acid sequence which is substantially homologous to the native sequence of an hCSF having at least one N-glycosylation site, wherein the mutant sequence comprises at least one amino acid substitution, deletion or insertion inactivating the N-glycosylation site.

DNA sequence encoding nonglycosylated analogs of human colony

Amplified expression of recombinant DNA products is achieved in hosts expressing proteases that cleave at multi-basic amino acid residues. To this end, cDNAs encoding granulocyte-macrophage colony stimulating factor (GM-CSF) are mutated such that one or both of the arginine residues at positions 23 and 24 of the protein product are deleted or replaced by non-basic amino acid residues. The GM-CSF analogs thus obtained maintain the activity of the wild-type protein.

Analogs of human granulocyte-macrophage colony stimulating factor

Amplified expression of recombinant DNA products is achieved in hosts expressing proteases that cleave at multi-basic amino acid residues. To this end, cDNAs encoding granulocyte-macrophage colony stimulating factor (GM-CSF) are mutated such that one or both of the arginine residues at positions 23 and 24 of the protein product are replaced by non-basic amino acid residues. The GM-CSF analogs thus obtained maintain the activity of the wild-type protein.

DNAs encoding analog GM-CSF molecules displaying resistance to proteases

Amplified expression of recombinant DNA products is achieved in hosts expressing protease that cleave at multi-basic amino acid residues. To this end, wild-type genes encoding the desired protein products are mutated by substituting codons or eliminating codons encoding multi-basic amino acid residues while maintaining the activity of the expressed protein product. Mutation of the desired gene can be conveniently carried out by site-specific in vitro mutagenisis.

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Seattle, WA, United States of America

Michael C Deeley