Merck Sharp + Dohme Corp.

Glycofi, Incorporated

Natarajan Sethuraman

Stefan Wildt

Bianka Prinz

Piotr Bobrowicz

Dongxing Zha

Robert Davidson

Terrance A Stadheim

Stephen Robin Hamilton

Juergen Nett

Byung-Kwon Choi

Michael A Meehl

Sujatha Gomathinayagam

Huijuan Li

Prinz Bianka

Robert C Davidson

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.

Surface display of whole antibodies in eukaryotes

Lower eukaryotic host cells have been engineered to produce glycoprotein having at least one terminal &#x3b1;-galactosyl epitope. The glycoproteins are useful for the production of highly antigenic glycoprotein compositions with advantages for the production of vaccines.

Yeast strain for the production of proteins with terminal alpha-1,3-linked galactose

Methods for increasing the yield and N-glycosylation site occupancy of paucimannose or complex N-glycans of recombinant glycoproteins produced in a recombinant host cell lacking dolichyl-P-Man:Man5GlcNAc2-PP-dolichyl alpha-1,3 mannosyltransferase (Alg3p) activity are disclosed. In particular, recombinant host cells are provided that comprise a disruption of the expression of an OS-9 family gene in the host cell. These recombinant host cells may then be used for producing recombinant glycoproteins. In further embodiments, the recombinant host cells further overexpress at least one heterologous single-subunit oligosaccharyltransferase, which in particular embodiments is capable of functionally suppressing the lethal phenotype of a mutation of at least one essential protein of the yeast oligosaccharyltransferase (OTase) complex.

Methods for increasing N-glycan occupancy and reducing production of hybrid N-glycans in pichia pastoris strains lacking ALG3 expression

Methods for producing proteins and glycoproteins inthat lack detectable cross binding activity to antibodies made against host cell antigens are described. In particular, methods are described wherein recombinantstrains that do not display a &#x3b2;-mannosyltransferase 2 activity with respect to an N-glycan or O-glycan and do not display at least one activity selected from a &#x3b2;-mannosyltransferase 1, 3, and 4 activity to produce recombinant proteins and glycoproteins. These recombinantstrains can produce proteins and glycoproteins that lack detectable &#x3b1;-mannosidase resistant &#x3b2;-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 inthat lack detectable cross binding activity to antibodies against host cell antigens.

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Described is a method for increasing the N-glycosylation site occupancy of a therapeutic glycoprotein produced in recombinant host cells modified as described herein and genetically engineered to express the glycoprotein compared to the N-glycosylation site occupancy of the therapeutic glycoprotein produced in a recombinant host cell not modified as described herein. In particular, the method provides recombinant host cells that overexpress a heterologous single-subunit oligosaccharyltransferase, which in particular embodiments is capable of functionally suppressing the lethal phenotype of a mutation of at least one essential protein of the yeast oligosaccharyltransferase (OTase) complex, for example, theSTT3D protein, in the presence of expression of the host cell genes encoding the endogenous OTase complex. The method is useful for both producing therapeutic glycoproteins with increased N-glycosylation site occupancy in lower eukaryote cells such as yeast and filamentous fungi and in higher eukaryote cells such as plant and insect cells and mammalian cells.

Methods and materials are provided for the production of compositions of erythropoietin protein, wherein said compositions comprise a pre-selected N-linked glycosylation pattern as the predominant N-glycoform.

Growing community of inventors

Hanover, NH, United States of America

Natarajan Sethuraman