In accordance with the present invention, CHO cells expressing a recombinant polypeptide of interest are grown in media where the amino acids, vitamins, phosphate, lipids and/or antioxidant optimization is utilized to manipulate and/or control the protein quality attributes of the polypeptides. Polypeptides expressed in accordance with the present invention may be advantageously used in the preparation of pharmaceutical compositions.

The disclosure provides synthetic (e.g. recombinant) pneumococcal saccharide comprising one or more repeat unit(s) .fwdarw.4)-β-D-Glcp-(1.fwdarw.3)-[[α-L-Rhap-(1.fwdarw.2)]-[Gro-(2.fwdarw.P.fwdarw.3)]-β-D-Galp-(1.fwdarw.4)]-β-L-Rhap-(1.fwdarw.. Also provided are conjugates comprising a .fwdarw.4)-β-D-Glcp-(1.fwdarw.3)-[[α-L-Rhap-(1.fwdarw.2)]-[Gro-(2.fwdarw.P.fwdarw.3)]-β-D-Galp-(1.fwdarw.4)]-β-L-Rhap-(1.fwdarw., immunogenic compositions, vaccines and their use in preventing or treating infection by Streptococcus pneumoniae.

The present invention relates to the finding of methods to shift the glycosylation profile of recombinant produced semm glycoproteins to the predominant bi-antennary form found in human plasma. This is accomplished by providing a mammalian cell line according to the invention with a series of gene disruptions and/or gene insertions that facilitate this shift.

The present invention provides inhibitors of fucosylation during protein expression from mammalian cells. The inhibitors are derived from rhamnose and act by inhibition of GDP-mannose 4,6-dehydratase (GMD). The invention further provides methods of making proteins with reduced level of fucosylation, such as antibodies and antibodies made by the methods of the present invention. Such hypofucosylated or nonfucosylated antibodies may find use, for example, in treatment of human disease in which is it therapeutically eneficial to direct antibody dependent cellular cytotoxicity (ADCC) mediated killing of cells expressing the antibody target on their surface, for example in depletion of Tregs in cancer patients using a hypofucosylated or nonfucosylated anti-CTLA-4 antibody.

Disclosed are a novel interferon lambda variant produced through structure-based glycoengineering and a method of producing the same. The novel interferon lambda variant and the method of producing the same exhibit remarkably improved production and yield in mammalian cell lines using structural information-based glycoengineering even through conventional purification protocols, and exhibit significantly improved therapeutic properties such as stability, half-life, and fraction of functional proteins during treatment, compared to wild-type interferon lambda. In addition, the novel interferon lambda variant and the method of producing the same according to the present invention have higher antiviral activity and interferon-stimulated gene (ISG)-inducing activity than wild-type interferon lambda, and thus are useful for the prevention and treatment of immune-related diseases such as cancer and autoimmune diseases as well as various viral infections such as infection with SARS-CoV-2 (COVID-19).

In certain embodiments, this disclosure provides a method of increasing production of a recombinant polypeptide of interest, comprising: a) seeding mammalian cells in a fed-batch production bioreactor at a viable cell density of at least 5106 viable cells/ml; and b) culturing the cells under optimized culture conditions to produce the recombinant polypeptide of interest at high titer.

The present invention is broadly concerned with new in vitro glycosylation methods that provide rational approaches for producing glycosylated proteins, and the use of glycosylated proteins. In more detail, the present invention comprises methods of glycosylating a starting protein having an amino sidechain with a nucleophilic moiety, comprising the step of reacting the protein with a carbohydrate having an oxazoline moiety on the reducing end thereof, to covalently bond the amino sidechain of the starting protein with the oxazoline moiety, wherein the glycosylated protein substantially retains the structure and function of the starting protein. Target proteins include oxidase, oxidoreductase and dehydrogenase enzymes. The glycosylated proteins advantageously have molecular weights of at least about 7500 Daltons. In a further embodiment, the present invention concerns the use of glycosylated proteins, fabricated by the methods disclosed herein, in the assembly of amperometric biosensors.

The present invention, in the field of bioengineering and biotechnology, relates to a method for preparing a recombinant antibody with a unique glycan profile produced by a genome-edited CHO host cell. Specifically, according to a method of the present invention, the TALEN technology is used to edit the FUT8 gene in CHO cells that have been adapted for serum-free suspension growth. The edited CHO host cells can produce recombinant antibodies with a unique glycan profile. The unique glycan profile can be characterized by non-fucosylated N-linked oligosaccharide chains of the antibodies, extremely low N-glycosylation heterogeneity and uniform carbohydrate chains. The antibody prepared by the method of the invention exhibit significantly increased ADCC and greater stability.

The present invention is broadly concerned with new in vitro glycosylation methods that provide rational approaches for producing glycosylated proteins, and the use of glycosylated proteins. In more detail, the present invention comprises methods of glycosylating a starting protein having an amino sidechain with a nucleophilic moiety, comprising the step of reacting the protein with a carbohydrate having an oxazoline moiety on the reducing end thereof, to covalently bond the amino sidechain of the starting protein with the oxazoline moiety, wherein the glycosylated protein substantially retains the structure and function of the starting protein. Target proteins include oxidase, oxidoreductase and dehydrogenase enzymes. The glycosylated proteins advantageously have molecular weights of at least about 7500 Daltons. In a further embodiment, the present invention concerns the use of glycosylated proteins, fabricated by the methods disclosed herein, in the assembly of amperometric biosensors.

The present invention relates to a mammalian cell comprising a gene encoding a polypeptide of interest, wherein the polypeptide of interest is expressed comprising one or more posttranslational modification patterns. These modifications are useful for example in glycoprotein production where the antibodies with the modifications have an enhanced antibody-dependent cell-mediated cytotoxicity (ADCC). The present invention also relates to methods for producing the glycoproteins and compositions comprising the glycoproteins, and their uses.