Chiron Corporation

Genetics Institute, Inc.

Other

Novartis Vaccines & Diagnostics, Gmbh

Patricia A Tekamp-Olson

Philip J Barr

Steven Rosenberg

Lawrence S Cousens

Jeffrey R Shuster

Randal J Kaufman

Anthony J Brake

James P Merryweather

Louise C Wasley

Polly A Wong

Pablo D Valenzuela

Guy T Mullenbach

Rae Lyn Burke

Mary Ellen Wernette-Hammond

Robert A Hallewell

Carol Ann Gallegos

Louise Wasley

Randal J 111 Marlborough Street Kaufman

Compositions and methods for expression of heterologous mammalian proteins and their secretion in the biologically active mature form using a yeast host cell as the expression system are provided. Compositions of the invention are nucleotide sequences encoding a signal peptide sequence for a yeast secreted protein, an optional leader peptide sequence for a yeast secreted protein, a native propeptide leader sequence for a mature protein of interest, and a sequence for the mature protein of interest, all operably linked to a yeast promoter. Each of these elements is associated with a processing site recognized in vivo by a yeast proteolytic enzyme. Any or all of these processing sites may be a preferred processing site that has been modified or synthetically derived for more efficient cleavage in vivo. The compositions are useful in methods for expression of heterologous mammalian proteins and their secretion in the biologically active mature form. Particularly, vectors comprising these nucleotide coding sequences can be used to transform a yeast host cell, which can then be cultured and screened for secretion of the biologically active mature protein of interest.

Method for expression of heterologous proteins in yeast

Inhibitors, including antibodies, of IL8 binding to its receptors, that interact with the amino-terminal extracellular domain of the IL8 receptor and which compete with IL8 for receptor-binding, are disclosed. The inhibitors are useful modulators of IL8 receptor-mediated biological activity.

IL8 inhibitors

A yeast &agr;-factor expression system is provided comprised of a truncated leader sequence, containing the &agr;-factor signal peptide and one glycosylation site, linked by a processing site to a non-yeast protein-encoding sequence.

Expression and secretion of heterologous proteins in yeast employing truncated alpha-factor leader sequences

Compositions and methods are provided for endopeptidase production and for enhanced efficiencies of processing heterologous precursor polypeptides to mature polypeptides, including proteins requiring gamma-carboxylation for biological activity. These compositions and methods utilize recombinant PACE, a mammalian endopeptidase that is specific for dibasic amino acid sites.

Expression of pace in host cells and methods of use thereof

Protein aggregates obtained by a process comprising culturing a host cell that expresses a desired protein in a medium comprising an effective amount of Cu.sup.++ and wherein the desired protein forms inclusion bodies in the host cell.

Enhanced purification and expression of insoluble recombinant proteins

Methods for the expression of mammalian MIP-1.alpha. and MIP-1.beta. are disclosed. The methods generally comprise introducing into a yeast cell, a DNA molecule capable of directing the expression and if desired the secretion of either MIP-1.alpha. or MIP-1.beta.. Methods for expression of constructs encoding both MIP-1.alpha. and MIP-1.beta. are also described. The MIP molecules so produced are biologically active.

Expression of macrophage inducible proteins (MIPS) in yeast cells

A yeast .alpha.-factor expression system is provided comprised of a truncated leader sequence, containing the .alpha.-factor signal peptide and one glycosylation site, linked by a processing site to a non-yeast protein-encoding sequence.

Expression and secretion of heterologous proteins in yeast employing

A method for isolating a desired protein which comprises: providing a host cell expressing the desired protein as an insoluble aggregate; culturing the host cell with an effective mount of Cu.sup.++ ; disrupting the host cell producing a lysate; incubating the insoluble fraction non-disulfide-bond reducing or non-copper competing chaotropic conditions to solubilize contaminants; separating the insoluble from the soluble fraction; and exposing the insoluble fraction to disulfide-bond reducing or copper competing chaotropic conditions to solubilize the desired protein.

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San Francisco, CA, United States of America

Patricia A Tekamp-Olson