Abstract: Methods for producing secreted receptor analogs and biologically active peptide dimers are disclosed. The methods for producing secreted receptor analogs and biologically active peptide dimers utilize a DNA sequence encoding a receptor analog or a peptide requiring dimerization for biological activity joined to a dimerizing protein. The receptor analog includes a ligand-binding domain. Polypeptides comprising essentially the extracellular domain of a human PDGF receptor fused to dimerizing proteins, the portion being capable of binding human PDGF or an isoform thereof, are also disclosed. The polypeptides may be used within methods for determining the presence of and for purifying human PDGF or isoforms thereof.

Abstract: Methods for producing secreted receptor analogs and biologically active peptide dimers are disclosed. The methods for producing secreted receptor analogs and biologically active peptide dimers utilize a DNA sequence encoding a receptor analog or a peptide requiring dimerization for biological activity joined to a dimerizing protein. The receptor analog includes a ligand-binding domain. Polypeptides comprising essentially the extracellular domain of a human PDGF receptor fused to dimerizing proteins, the portion being capable of binding human PDGF or an isoform thereof, are also disclosed. The polypeptides may be used within methods for determining the presence of and for purifying human PDGF or isoforms thereof.

Abstract: Methods for producing secreted receptor analogs and biologically active peptide dimers are disclosed. The methods for producing secreted receptor analogs and biologically active peptide dimers utilize a DNA sequence encoding a receptor analog or a peptide requiring dimerization for biological activity joined to a dimerizing protein. The receptor analog includes a ligand-binding domain. Polypeptides comprising essentially the extracellular domain of a human PDGF receptor fused to dimerizing proteins, the portion being capable of binding human PDGF or an isoform thereof, are also disclosed. The polypeptides may be used within methods for determining the presence of and for purifying human PDGF or isoforms thereof.

Abstract: Proteinase inhibitors comprising a Kunitz domain are disclosed. The Kunitz domain comprises a sequence of amino acid residues as shown in SEQ ID NO:5, wherein the sequence is at least 90% identical to SEQ ID NO:2. Also disclosed are methods for making the proteinase inhibitors, and expression vectors and cultured cells that are useful within the methods. The proteinase inhibitors may be used as components of cell culture media, in protein purification, and in certain therapeutic and diagnostic applications.

Abstract: Methods for preparing double stranded, circular DNA molecules comprising a region encoding a protein of interest are disclosed. One or more donor DNA fragments encoding the protein of interest are combined with an acceptor plasmid, a first DNA linker, and a second DNA linker in a Saccharomyces cerevisiae host cell whereby the donor DNA fragment is joined to the acceptor plasmid by homologous recombination of the donor DNA, acceptor plasmid, and linkers to form the closed, circular plasmid.

Abstract: Methods for altering a selected chromosomal locus in P. methanolica cells and cells comprising such altered loci are disclosed. A linear DNA construct comprising (i) a segment comprising a portion of the target locus in which at least one nucleotide pair is altered and (ii) a selectable marker that complements adenine auxotrophy is introduced into cells auxotrophic for adenine. The cells are cultured under selective conditions, and cells in which the linear DNA construct has been chromosomally integrated by homologous recombination are identified. The cells are then cultured under conditions whereby cells auxotrophic for adenine can be identified, and a subset of such cells in which the altered locus has been chromosomally integrated are identified.

Abstract: A novel serine protease is disclosed. The protease comprises a sequence of amino acid residues that is at least 95% identical to SEQ ID NO:2 from Ile, residue 111, through Asn, residue 373. Also disclosed are polynucleotide molecules encoding the protease, expression vectors containg the polynucleotides, cultured cells containing the expression vectors, and methods of making the protease. The protease can be used, inter alia, within industrial processes to degrade unwanted proteins or alter the characteristics of protein-containing compositions.

Abstract: Protease-deficient strains of the methylotrophic yeast Pichia methanolica and materials and methods for generating such strains are disclosed. The strains have a functional deficiency in a vacuolar protease, such as proteinase A or proteinase B. The strains are useful as hosts for the expression of heterologous genes encoding proteins of commercial or other interest.

Abstract: Proteinase inhibitors comprising a Kunitz domain are disclosed. The Kunitz domain comprises a sequence of amino acid residues as shown in SEQ ID NO:5, wherein the sequence is at least 90% identical to SEQ ID NO:2. Also disclosed are methods for making the proteinase inhibitors, and expression vectors and cultured cells that are useful within the methods. The proteinase inhibitors may be used as components of cell culture media, in protein purification, and in certain therapeutic and diagnostic applications.

Abstract: Novel receptor polypeptides, polynucleotides encoding the polypeptides, and related compositions and methods are disclosed. The polypeptides comprise an extracellular ligand-binding domain of a cell-surface receptor that is expressed at high levels in lymphoid tissue, including B-cells and T-cels. The polypeptides may be used within methods for detecting ligands that stimulate the proliferation and/or development of lymphoid and myeloid cells in vitro and in vivo. Ligand-binding receptor polypeptides can also be used to block ligand activity in vitro and in vivo.

Abstract: Dimeric proteins having substantially the same biological activity as PDGF are disclosed. More specifically, the protein may have two substantially identical polypeptide chains, each of the chains being substantially homologous to the A-chain of PDGF. Alternatively, the protein may have two polypeptide chains that are substantially identical to the A-chain of PDGF. In addition, proteins comprising polypeptides that are variants or derivatives of the A-chain of PDGF are also disclosed. Therapeutic compositions containing these proteins and methods for enhancing the wound-healing process in warm-blooded animals are also disclosed.

Abstract: Pichia methanolica cells in which an alcohol oxidase gene has been disrupted are disclosed. The cells may also be deficient in vacuolar protease activity. The cells are useful as hosts in recombinant protein production methods.

Abstract: Purified mammalian thrombopoietin proteins and methods of making them are disclosed. The proteins are characterized by a M.sub.r =70,000.+-.10,000 daltons as determined by SDS-polyacrylamide gel electrophoresis under denaturing conditions and are at least 90% pure with respect to contaminating proteins. The proteins can be prepared by a method in which thrombopoietin is adsorbed to and eluted from a polypeptide comprising a ligand-binding domain of an MPL receptor, then the eluted thrombopoietin is fractionated by anion exchange chromatography.

Abstract: Methods for inhibiting intimal hyperplasia in the vasculature of mammals, including primates, are disclosed. The methods comprise administering to the mammal an anti-PDGF receptor antibody, such as an anti-PDGF-alpha receptor antibody or an anti-PDGF-beta receptor antibody. The methods are useful in reducing intimal hyperplasia due to, for example, vascular injuries resulting from angioplasty, endarterectomy, reduction atherectomy or anastomosis of a vascular graft. The anti-PDGF receptor antibodies may optionally be administered coordinately with heparin, whereby the coordinately administered antibody and heparin are combinatorially effective in inhibiting intimal hyperplasia.

Abstract: DNA constructs, Pichia methanolica cells, and methods for producing polypeptides using the DNA constructs and cells are disclosed. The DNA constructs comprise a transcription promoter of a Pichia methanolica gene, a DNA segment encoding a polypeptide heterologous to P. methanolica, a transcription terminator of a P. methanolica gene, and a selectable marker. P. methanolica cells containing the DNA constructs are used within methods for producing polypeptides heterologous to P. methanolica. DNA molecules comprising transcription promoters of P. methanolica genes are also disclosed.

Abstract: Polypeptides of from about 9-120 amino acid residues comprising a truncated segment of fibrinogen which may be flanked by elastomeric peptides, wherein the polypeptides are cross-linkable by a transglutaminase, homo- and copolymers containing such polypeptides are disclosed. The homo- and co-polymers disclosed herein are useful in tissue sealant and wound healing formulations.

Abstract: Novel receptor polypeptides, polynucleotides encoding the polypeptides, and related compositions and methods are disclosed. The polypeptides comprise an extracellular ligand-binding domain of a cell-surface receptor that is expressed at high levels in lymphoid tissue, including B-cells and T-cells. The polypeptides may be used within methods for detecting ligands that stimulate the proliferation and/or development of lymphoid and myeloid cells in vitro and in vivo. Ligand-binding receptor polypeptides can also be used to block ligand activity in vitro and in vivo.

Abstract: The present invention provides isolated DNA molecules comprising a DNA segment encoding novel human Kunitz-type inhibitors. Also provided are DNA constructs comprising a first DNA segment encoding a novel human Kunitz-type inhibitor wherein first DNA segment is operably linked to additional DNA segments required for the expression for the first DNA segment, as well as host cells containing such DNA constructs and methods for producing proteins from the host cells.

Abstract: DNA constructs, Pichia methanolica cells, and methods for producing polypeptides using the DNA constructs and cells are disclosed. The DNA constructs comprise a transcription promoter of a Pichia methanolica gene, a DNA segment encoding a polypeptide heterologous to P. methanolica, a transcription terminator of a P. methanolica gene, and a selectable marker. P. methanolica cells containing the DNA constructs are used within methods for producing polypeptides heterologous to P. methanolica. DNA molecules comprising transcription promoters of P. methanolica genes are also disclosed.

Abstract: Methods are provided for producing protein products in host cells and for selecting transformed cells comprising the step of transforming the host cell with a DNA molecule comprising a gene which complements a deficiency in the host cell. The host cell is s strain having a deficiency in a function necessary for normal cell growth. The gene in the DNA molecule, such as a plasmid, which complements the deficiency serves as a selection marker whereby the growth conditions for selection may comprise a conventional complex medium.