Disclosed are new recombinant isoforms of human-like lubricin or PRG4 glycoprotein having outstanding lubrication properties and a novel glycosylation pattern, and methods for their manufacture at high levels enabling commercial production.

The invention relates to compositions including polynucleotides encoding polypeptides which have been chemically modified by replacing the uridines with 1-methyl-pseudouridine to improve one or more of the stability and/or clearance in tissues, receptor uptake and/or kinetics, cellular access by the compositions, engagement with translational machinery, mRNA half-life, translation efficiency, immune evasion, protein production capacity, secretion efficiency, accessibility to circulation, protein half-life and/or modulation of a cell's status, function, and/or activity.

The present invention relates to methods and compositions for modulating mannosylation profile of recombinant proteins expressed by mammalian host cells during the cell culture process, using a polyether ionophore.

The invention concerns fusion proteins, wherein two endoglycosidases are fused, possibly via a linker. The fusion enzymes according to the invention have structure (1): EndoX-(L).sub.p-EndoY (1), wherein EndoX is an endoglycosidase, EndoY is an endoglycosidase distinct from EndoX, L is a linker and p is 0 or 1. Such fusion enzymes capable of trimming glycoproteins comprising at least two distinct glycoforms in a single step. The invention further concerns the use of the fusion enzyme according to the invention for trimming glycoproteins. In another aspect, the invention relates to the process of production of the fusion enzyme. In a further aspect, the inventions concerns a process for trimming glycoproteins, comprising trimming the glycoprotein with a fusion enzyme according to the invention, to obtain a trimmed glycoprotein.

The present invention relates to cell culture media comprising 5-Thio-L-fucose and the use of 5-Thio-L-fucose for modulating the glycosylation profile of antibodies or other Fc containing proteins and thereby modulating the binding affinity of said proteins.

An isolated nucleic acid encoding an FX protein having a serine at position 79, a lysine at position 90, a leucine at position 136, an arginine at position 211, a serine at position 289, and a combination thereof is provided. Cells having a gene encoding a modified FX protein are provided, wherein the cells exhibit a reduced ability to fucosylate a glycoprotein at a first temperature, but exhibit the ability to fucosylate the glycoprotein at a second temperature. Methods and compositions for making glycoproteins with reduced fucosylation are provided.

Royal jelly honey produced by nurse bees, honey and milk has great values in human health and/or beauty. This invention aims to produce royal jelly honey substitute, honey substitute, animal free milk without the need for bees or cows or goats. First, recombinant major royal jelly proteins (MRJPs), defensin-1, and milk proteins are produced by microbes (such as bacteria, yeasts, or fungi) using precision fermentation. The MRJPs, defensin-1 or milk proteins are then formulated with vitamins, water, minerals, fats, salt and/or amino acids to produce a bee-free royal jelly honey substitute, bee-free honey substitute or animal free milk. MRJPs can also be formulated with vitamins, water, minerals, fats, salt, amino acids and recombinant milk proteins to produce animal-free milk or milk substitutes with supplemental MRJPs. Plant-based milks can also be supplemented with MRJPs and recombinant milk proteins.

Materials and methods are provided for making plants (e.g., Nicotiana varieties) that are suitable for producing therapeutic polypeptides suitable for administration to humans and animals, particularly by making TAL effector endonuclease-induced mutations in genes encoding xylosyltransferases and fucosyltransferases.

The present invention relates to a method for increasing the expression and/or promoting correct folding of a heterologous polypeptide of interest in a plant cell, comprising co-expressing the heterologous polypeptide of interest with a polypeptide encoding a mammalian chaperone protein. The invention also relates to plant cells and plants, which either transiently or stably, co-express the heterologous polypeptide of interest and the chaperone protein.

A method of modifying a glycosylation pattern of a polypeptide-of-interest in a plant or plant cell is provided. The method comprising expressing in a plant or plant cell transformed to express at least one glycosidase in a subcellular compartment, a nucleic acid sequence encoding the polypeptide-of-interest, such that the at least one glycosidase and the polypeptide-of-interest are co-localized to the subcellular compartment of the plant or plant cell, thereby modifying the glycosylation pattern of the polypeptide-of-interest in the plant or plant cell.