Zymogenetics, Inc.

University of Washington

University of Texas System

Zymogenetrics, Inc.

Frederick S Hagen

Eileen R Mulvihill

Khaled M Houamed

Wolfhard Almers

Mark J Murray

Kathleen L Berkner

Sharon J Busby

Kenneth Kaushansky

Margaret Y Insley

Richard G Woodbury

Charles L Gray

Donald C Foster

Joseph Leon Kuijper

Francis J Grant

Richard D Holly

Si Lok

Andrzej Z Sledziewski

Patrick J O'Hara

Catherine E Lofton-Day

Robert S Munford

Anur A Kumar

Pieter J Oort

Methods are disclosed for producing proteins having biological activity for blood coagulation mediated by Factor VIIa or Factor IX. The proteins are produced by mammalian host cells which have been stably transfected with a DNA construct containing a nucleotide sequence which codes at least partially for either Factor VII or Factor IX. The nucleotide sequence comprises a first nucleotide sequence encoding a calcium binding domain, joined to a second nucleotide sequence positioned downstream of the first sequence. The second sequence encodes a catalytic domain for the serine protease activity of either Factor VIIa or Factor IX. The joined sequences code for proteins having substantially the same biological activity for blood coagulation as either Factor VIIa or Factor IX.

Expression of factor VII and IX activities in mammalian cells

Methods for stimulating granulocyte/macrophage lineage cells using thrombopoiet in are provided. The methods provided may be used to stimulate granulocyte/macrophage progenitors and neutrophils in bone marrow and peripheral blood cells and in vitro and in vivo. In addition, methods for treatment of neutropenia in patients are disclosed.

Methods for stimulating granulocyte/macrophage lineage using

Mammalian G protein coupled glutamate receptors are identified, isolated and purified. The receptors have been cloned, sequenced and expressed by recombinant means. The receptors and antibodies thereby may be used to identify agonists and antagonists of G protein coupled glutamate receptor mediated neuronal excitation, as well as in methods of diagnosis.

G protein coupled glutamate receptors

Method for identifying a G protein coupled glutamate receptor agonist

Antibodies to G protein coupled glutamate receptors

Novel proteins possessing substantially the same biological activity as human GM-CSF, as well as proteins having a higher specific activity than native human GM-CSF, are disclosed. The proteins may (a) be unglycosylated; (b) lack N-linked glycosylation but have O-linked glycosylation; or (c) lack one of the two N-linked carbohydrate chains characteristic of native human GM-CSF. The proteins are produced using recombinant DNA techniques in eucaryotic host cells, such as mammalian, yeast, and filamentous fungal host cells. Pharmaceutical compositions including an effective amount of one of the proteins of the invention and a physiologically acceptable carrier or diluent are also disclosed.

Colony-stimulating factor derivatives

Method for producing a mammalian G protein coupled glutamate receptor

Methods are disclosed for producing acyloxyacyl hydrolase. The protein is produced from eukaryotic host cells transformed or transfected with DNA construct(s) containing information necessary to direct the expression of acyloxyacyl hydrolase. The DNA constructs generally include the following operably linked elements: a transcriptional promoter; DNA sequence encoding acyloxyacyl hydrolase, the small subunit of acyloxyacyl hydrolase or the large subunit of acyloxyacyl hydrolase; and a transcriptional terminator. In addition, isolated DNA sequences encoding acyloxyacyl hydrolase and isolated DNA sequences encoding the small or large subunit of acyloxyacyl hydrolase are disclosed.

Method for producing acyloxyacyl hydrolase

Methods for purifying factor VIII:C, von Willebrand factor (vWF) or complexes thereof from heterogeneous biological fluids are disclosed. The methods utilize a binding peptide, specific to either factor VIII:C or vWF, bound to an insoluble matrix. Peptides suitable for use within the methods are also disclosed.

Method for purifying factor VIII:C, von Willebrand factor and complexes

Methods for cloning cDNA and producing mRNA from the cloned cDNA are disclosed. The cDNA is cloned in one orientation through the use of a vector containing a directional cloning site and through the use of primer-adapter sequences complementary to portions of the directional cloning site. The vector may also contain a promoter, one or more terminators and a polyadenylation signal to facilitate transcription of the cloned cDNA and translation of the mRNA.

Directional cloning

Methods are disclosed for producing proteins having biological activity for blood coagulation mediated by Factor VIIa. The proteins are produced by mammalian host cells which have been stably transfected with a DNA construct containing a nucleotide sequence which codes at least partially for either Factor VII. The nucleotide sequence comprises a first nucleotide sequence encoding a calcium binding domain, joined to a second nucleotide sequence positioned downstream of the first sequence. In particular, the first nucleotide sequence may be derived from a genomic clone or cDNA clone of Factor VII. The second sequence encodes a catalytic domain for the serine protease activity of Factor VIIA. The joined sequences code for proteins having substantially the same biological activity for blood coagulation as Factor VIIa.

Expression of factor VII activity in mammalian cells

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Seattle, WA, United States of America

Frederick S Hagen