Genetics Institute, Inc.

Novartis Corporation

Chiron Corporation

Genetics Institute, Inc. - Legal Affairs

Randal J Kaufman

Steven C Clark

Gordon G Wong

Debra D Pittman

Louise C Wasley

Philip J Barr

Patricia A Tekamp-Olson

Anthony J Brake

Polly A Wong

Elizabeth A Wang

John J Toole, Jr

John Martin Wozney

S Robert Adamson

Charles B Shoemaker

Andrew Dorner

Alnawaz Rehemtulla

Gordon C Wong

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

A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises: preparing RNA from a cell that produces CSF; preparing polyadenylated messenger RNA from said RNA; preparing single stranded cDNA from said messenger RNA; converting the single stranded cDNA to double stranded cDNA; inserting the double stranded cDNA into transformation vectors and transforming bacteria with said vector to form colonies; picking pools of 200 to 500 colonies each and isolating plasmid DNA from each pool; transfecting the plasmid DNA into suitable host cells for expressing CSF protein; culturing the transfected cells and assaying the supernatant for CSF activity; and selecting CSF positive pools and screening the colonies used to make the pool to identify a colony having CSF activity. Also described are a cDNA coding for a protein having CSF activity (i.e. CSF/cDNA), a microorganism or cell line transformed with a recombinant vector containing such CSF/cDNA, and a method for producing CSF protein by expressing said CSF/cDNA by culturing a microorganism or cell line.

Recombinant primate granulocyte macrophage-colony stimulating factor

A method for preparing and isolating a transformation vector containing CSF/cDNA is described. The method comprises:

DNA encoding GM-CSF and a method of producing GM-CSF protein

Recombinant human granulocyte-macrophage-colony stimulating factor

Method of treating myelo-suppression with GM-CSF

Provided are porcine factor VIII nucleotide sequences and hybrid sequences encoding human/porcine chimeric factor VIII-type procoagulant proteins. DNAs encoding such chimeric factor VIII-type procoagulant proteins, pharmaceutical compositions containing such chimeric factor VIII-type procoagulant proteins, and methods of treating hemophilia using such chimeric factor VIII-type procoagulant proteins are also provided.

Chimeric procoagulant proteins

Expression of PACE in host cells and methods of use thereof

This invention relates to recombinant Factor VIII:c variants, methods to produce the variants and pharmaceutical compositions containing same. The variants of this invention are characterized by modification of one or more specific proteolytic cleavage sites encompassing the arginine residues at positions 226, 336, 562, 740, 776, 1313, 1648, or 1721. The variants possess similar procoagulant activity to that of human Factor VIII:c.

Procoagulant proteins

This invention relates to recombinant Factor VIII:c variants, methods to produce the variants and pharmaceutical compositions containing same.

Human factor VIII:c muteins

An improved method for producing Factor VIII:c-type proteins is disclosed which involves culturing mammalian cells which are capable of expressing the protein. In accordance with this invention the cells are cultured in a medium containing an effective amount of a substance comprising (a) von Willebrand Factor-type protein, (b) a phospholipid or phospholipid mixture, or a mixture of (a) and (b).

Method for producing factor VIII:C-type proteins

A transformation vector containing a heterologous product gene and two or more different heterologous selectable amplifiable marker genes is described. Preferably, the marker genes form a polycistronic transcription unit. Also described is a method for obtaining high level expression of a desired protein by culturing eukaryotic cells containing the vectors of the invention.

Multiply-amplifiable vectors for high level expression of exogenuos DNA

An improved method for producing Factor VIII:c is disclosed. The method involves culturing mammalian cells which contain DNA encoding Factor VIII:c and which are capable of expressing Factor VIII:c. In accordance with this invention the cells are cultured in a medium containing an effective amount of a Factor VIII:c-stabilizing substance comprising (a) von Willebrand Factor (VWF), (b) a phospholipid or phospholipid mixture, or a mixture of (a) and (b).

Method for producing factor VIII:C and analogs

Eucaryotic cells cotransformed with product and selection genes yield considerably greater quantities of product after a novel subcloning strategy is employed: Transformants are identified for product yield, cultured under selection pressure and the progeny screened for product yield. Novel transformation vectors contain directly ligated selection and product genes and/or eucaryotic promoters.

Method for making tissue plasminogen activator

Hybrid procoagulant proteins are disclosed which contain peptide sequences of human blood coagulation factors V and VIII. DNA molecules encoding these proteins and materials and methods for expressing them are also disclosed. Preferably, peptide sequence in the B domain of Factor VIII is replaced with peptide sequence derived from human Factor V.

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Boston, MA, United States of America

Randal J Kaufman