Patents by Inventor Piotr Bobrowicz

Piotr Bobrowicz has filed for patents to protect the following inventions. This listing includes patent applications that are pending as well as patents that have already been granted by the United States Patent and Trademark Office (USPTO).

  • Patent number: 8697394
    Abstract: The present invention relates to eukaryotic host cells, especially lower eukaryotic host cells, having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar and sugar nucleotide transporters to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells exhibit GnTIII, GnTIV, GnTV, GnT VI or GnTIX activity, which produce bisected and/or multiantennary N-glycan structures and may be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar, sugar nucleotide transporters, to yield human-like glycoproteins.
    Type: Grant
    Filed: February 20, 2004
    Date of Patent: April 15, 2014
    Assignee: Glycofi, Inc.
    Inventors: Piotr Bobrowicz, Stephen R. Hamilton, Tillman U. Gerngross, Stefan Wildt, Byung-Kwon Choi, Juergen Hermann Nett, Robert C. Davidson
  • Publication number: 20130295608
    Abstract: Lower eukaryotic host cells have been recombinantly engineered to produce glycoprotein having human-like O-glycosylation. The glycoproteins are useful for the production of glycoprotein compositions with advantages for the production of human therapeutics.
    Type: Application
    Filed: February 20, 2012
    Publication date: November 7, 2013
    Inventors: Piotr Bobrowicz, William J. Cook, Stephen Hamilton, Juergen Nett, Terrance A. Stadheim, Stefan Wildt
  • Publication number: 20130217067
    Abstract: The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities such as those involved in glycosylation to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified oligosaccharides are created or selected. N-glycans made in the engineered host cells have a Man5GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g.
    Type: Application
    Filed: February 29, 2012
    Publication date: August 22, 2013
    Applicant: GlycoFi, Inc.
    Inventors: TILLMAN U. GERNGROSS, Stefan Wildt, Byung-kwon Choi, Juergen Hermann Nett, Piotr Bobrowicz, Stephen R. Hamilton, Robert C. Davidson
  • Patent number: 8501438
    Abstract: A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more ?-1,2-mannosidases.
    Type: Grant
    Filed: May 31, 2012
    Date of Patent: August 6, 2013
    Assignee: GlycoFi, Inc.
    Inventors: Piotr Bobrowicz, W. James Cook, Warren Kett
  • Patent number: 8445227
    Abstract: The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified lipid-linked oligosaccharides are created or selected. N-glycans made in the engineered host cells exhibit GnTIII activity, which produce bisected N-glycan structures and may be modified further by heterologous expression of one or more enzymes, e.g., glycosyltransferases, sugar transporters and mannosidases, to yield human-like glycoproteins. For the production of therapeutic proteins, this method may be adapted to engineer cell lines in which any desired glycosylation structure may be obtained.
    Type: Grant
    Filed: August 13, 2009
    Date of Patent: May 21, 2013
    Assignee: Merck Sharp & Dohme
    Inventors: Piotr Bobrowicz, Stephen R. Hamilton, Tillman U. Gerngross, Stefan Wildt, Byung-Kwon Choi, Juergen Hermann Nett, Robert C. Davidson
  • Publication number: 20120322101
    Abstract: A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more ?-1,2-mannosidases.
    Type: Application
    Filed: June 22, 2012
    Publication date: December 20, 2012
    Applicant: GLYCOFI, INC.
    Inventors: Piotr Bobrowicz, W. James Cook, Warren Kett
  • Publication number: 20120237973
    Abstract: A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more ?-1,2-mannosidases.
    Type: Application
    Filed: May 31, 2012
    Publication date: September 20, 2012
    Applicant: GLYCOFI, INC.
    Inventors: Piotr Bobrowicz, W. James Cook, Warren Kett
  • Publication number: 20120232007
    Abstract: Methods for producing proteins and glycoproteins in Pichia pastoris that lack detectable cross binding activity to antibodies made against host cell antigens are described. In particular, methods are described wherein recombinant Pichia pastoris strains that do not display a ?-mannosyltransferase 2 activity with respect to an N-glycan or O-glycan and do not display at least one activity selected from a ?-mannosyltransferase 1, 3, and 4 activity to produce recombinant proteins and glycoproteins. These recombinant Pichia pastoris strains can produce proteins and glycoproteins that lack detectable ?-mannosidase resistant ?-mannose residues thereon and thus, lack cross binding activity to antibodies against host cell antigens. Further described are methods for producing bi-sialylated human erythropoietin in Pichia pastoris that lack detectable cross binding activity to antibodies against host cell antigens.
    Type: Application
    Filed: October 11, 2010
    Publication date: September 13, 2012
    Applicant: MERCK SHARP & DOHME CORP
    Inventors: Piotr Bobrowicz, Sujatha Gomathinayagam, Stephen Hamilton, Huijuan Li, Natarajan Sethuraman, Terrance A. Stadheim, Stefan Wildt
  • Publication number: 20120213728
    Abstract: Compositions comprising granulocyte-colony stimulating factor (GCSF) produced in a strain of Pichia pastoris glycoengineered to produce a GCSF wherein greater than 18% of the molecules comprise an 0-glycan with one mannose per (0-glycan is described. In particular aspects, the GCSF is PEGylated at the JV-terminus.
    Type: Application
    Filed: October 25, 2010
    Publication date: August 23, 2012
    Applicant: MERCK
    Inventors: Michael Meehl, Sandra Rios, Sujatha Gomathinayagam, Huijuan Li, Piotr Bobrowicz
  • Patent number: 8206949
    Abstract: A method is described for producing protein compositions having reduced amounts of O-linked glycosylation. The method includes producing the protein in cells cultured in the presence of an inhibitor of Pmt-mediated O-linked glycosylation and/or in the presence of one or more ?-1,2-mannosidases.
    Type: Grant
    Filed: November 10, 2006
    Date of Patent: June 26, 2012
    Assignee: GlycoFi, Inc.
    Inventors: Piotr Bobrowicz, W. James Cook, Warren Kett
  • Patent number: 8158384
    Abstract: The present invention provides methods to reduce or eliminate ?-mannosidase resistant glycans on glycoproteins in yeast. The reduction or elimination of ?-mannosidase resistant glycans on glycoproteins results from the disruption of the newly isolated P. pastoris AMR2 gene encoding ?1,2-mannosyltransferase. The present invention also discloses novel genes, polypeptides, antibodies, vectors and host cells relating to ?-mannosidase resistance on glycans.
    Type: Grant
    Filed: July 8, 2008
    Date of Patent: April 17, 2012
    Assignee: Glycofi, Inc.
    Inventor: Piotr Bobrowicz
  • Publication number: 20120052530
    Abstract: The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities such as those involved in glycosylation to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified oligosaccharides are created or selected. N-glycans made in the engineered host cells have a Man5GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g.
    Type: Application
    Filed: June 9, 2011
    Publication date: March 1, 2012
    Applicant: GlycoFi, Inc.
    Inventors: TILLMAN U. GERNGROSS, Stefan Wildt, Byung-kwon Choi, Juergen Hermann Nett, Piotr Bobrowicz, Stephen R. Hamilton, Robert C. Davidson
  • Publication number: 20120021948
    Abstract: Methods for display of recombinant whole immunoglobulins or immunoglobulin libraries on the surface of eukaryote host cells, including yeast and filamentous fungi, are described. The methods are useful for screening libraries of recombinant immunoglobulins in eukaryote host cells to identify immunoglobulins that are specific for an antigen of interest.
    Type: Application
    Filed: October 3, 2011
    Publication date: January 26, 2012
    Applicant: Merck Sharp & Dohme Corp.
    Inventors: Bianka Prinz, Natarajan Sethuraman, Dongxing Zha, Stefan Wildt, Piotr Bobrowicz
  • Publication number: 20120003695
    Abstract: Lower eukaryotic cells such as Pichia pastoris that normally cannot use galactose as a carbon source but which have been genetically engineered according to the methods herein to use galactose as a sole source of carbon are described. The cells are genetically engineered to express several of the enzymes comprising the Leloir pathway. In particular, the cells are genetically engineered to express a galactokinase, a UDP-galactose-C4-epimerase, and a galactose-1-phosphate uridyltransferase, and optionally a galactose permease. In addition, a method is provided for improving the yield of glycoproteins that have galactose-terminated or -containing N-glycans in cells that have been genetically engineered to produce glycoproteins with N-glycans having galactose residues but which normally lack the enzymes comprising the Leloir pathway comprising transforming the cells with one or more nucleic acid molecules encoding a galactokinase, a UDP-galactose-C4-epimerase, and a galactose-1-phosphate uridyltransferase.
    Type: Application
    Filed: February 24, 2010
    Publication date: January 5, 2012
    Inventors: Robert C. Davidson, Piotr Bobrowicz, Dongxing Zha
  • Publication number: 20110312032
    Abstract: Lower eukaryote host cells in which the function of at least one endogenous gene encoding a chaperone protein, such as a Protein Disulphide Isomerase (PDI), has been reduced or eliminated and at least one mammalian homolog of the chaperone protein is expressed are described. In particular aspects, the host cells further include a deletion or disruption of one or more O-protein mannosyltransferase genes, and/or overexpression of an endogenous or exogenous Ca2+ATPase. These host cells are useful for producing recombinant glycoproteins in large amounts and for producing recombinant glycoproteins that have reduced O-glycosylation.
    Type: Application
    Filed: August 11, 2011
    Publication date: December 22, 2011
    Inventors: Byung-Kwon Choi, Piotr Bobrowicz, W. James Cook, Elena E. Brevnova
  • Patent number: 8067339
    Abstract: Methods for display of recombinant whole immunoglobulins or immunoglobulin libraries on the surface of eukaryote host cells, including yeast and filamentous fungi, are described. The methods are useful for screening libraries of recombinant immunoglobulins in eukaryote host cells to identify immunoglobulins that are specific for an antigen of interest.
    Type: Grant
    Filed: June 23, 2009
    Date of Patent: November 29, 2011
    Assignee: Merck Sharp & Dohme Corp.
    Inventors: Bianka Prinz, Natarajan Sethuraman, Dongxing Zha, Stefan Wildt, Piotr Bobrowicz
  • Patent number: 8067551
    Abstract: The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities such as those involved in glycosylation to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified oligosaccharides are created or selected. N-glycans made in the engineered host cells have a Man5GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g.
    Type: Grant
    Filed: November 7, 2008
    Date of Patent: November 29, 2011
    Assignee: Glycofi, Inc.
    Inventors: Tillman U. Gerngross, Stefan Wildt, Byung-Kwon Choi, Juergen Hermann Nett, Piotr Bobrowicz, Stephen R Hamilton, Robert C. Davidson
  • Patent number: 7935513
    Abstract: The present invention relates to eukaryotic host cells having modified oligosaccharides which may be modified further by heterologous expression of a set of glycosyltransferases, sugar transporters and mannosidases to become host-strains for the production of mammalian, e.g., human therapeutic glycoproteins. The invention provides nucleic acid molecules and combinatorial libraries which can be used to successfully target and express mammalian enzymatic activities such as those involved in glycosylation to intracellular compartments in a eukaryotic host cell. The process provides an engineered host cell which can be used to express and target any desirable gene(s) involved in glycosylation. Host cells with modified oligosaccharides are created or selected. N-glycans made in the engineered host cells have a Man5GlcNAc2 core structure which may then be modified further by heterologous expression of one or more enzymes, e.g.
    Type: Grant
    Filed: June 5, 2008
    Date of Patent: May 3, 2011
    Assignee: Glycofi, Inc.
    Inventors: Tillman U. Gerngross, Stefan Wildt, Byung-Kwon Choi, Juergen Hermann Nett, Piotr Bobrowicz, Stephen R. Hamilton, Robert C. Davidson
  • Publication number: 20110053214
    Abstract: The present invention provides a novel lower eukaryotic host cell producing human-like glycoproteins characterized as having a terminal ?-galactose residue and essentially lacking fucose and sialic acid residues. The present invention also provides a method for catalyzing the transfer of a galactose residue from UDP-galactose onto an acceptor substrate in a recombinant lower eukaryotic host cell, which can be used as a therapeutic glycoprotein.
    Type: Application
    Filed: July 21, 2010
    Publication date: March 3, 2011
    Applicant: GLYCOFI, INC.
    Inventors: Robert Collier Davidson, Tillman Ulf Gerngross, Stefan Wildt, Byung-Kwon Choi, Juergen Hermann Nett, Piotr Bobrowicz, Stephen Robin Hamilton
  • Publication number: 20100311122
    Abstract: Lower eukaryote host cells in which the function of at least one endogenous gene encoding a chaperone protein, such as a Protein Disulphide Isomerase (PDI), has been reduced or eliminated and at least one mammalian homolog of the chaperone protein is expressed are described. In particular aspects, the host cells further include a deletion or disruption of one or more O-protein mannosyltransferase genes, and/or overexpression of an endogenous or exogenous Ca2+ ATPase. These host cells are useful for producing recombinant glycoproteins in large amounts and for producing recombinant glycoproteins that have reduced O-glycosylation.
    Type: Application
    Filed: February 9, 2009
    Publication date: December 9, 2010
    Inventors: Byung-Kwon Choi, Piotr Bobrowicz, W. James Cook