The present invention comprises a method of concentrating a composition comprising a polypeptide of interest and the use of such concentrated composition for the treatment of diseases in mammals, in particular by subcutaneous injection.

The invention features a method of producing a high mannose glucocerebrosidase (hmGCB) which includes: providing a cell which is capable of expressing glucocerebrosidase (GCB), and allowing production of GCB having a precursor oligosaccharide under conditions which prevent the removal of at least one mannose residue distal to the pentasaccharide core of the precursor oligosaccharide of GCB, to thereby produce an hmGCB preparation. Preferably, the condition which prevents the removal of at least one mannose residue distal to the pentasaccharide core is inhibition of a class 1 processing mannosidase and/or a class 2 processing mannosidase. The invention also features an hmGCB preparation and methods of using an hmGCB preparation.

Described herein are methods and genetically engineered cells useful for producing an altered N-glycosylation form of a target molecule. Also described are methods and molecules with altered N-glycosylation useful for treating a variety of disorders such as metabolic disorders.

The present invention relates to a process for purification of recombinant alpha-mannosidase, a process for production of alpha-mannosidase, a composition comprising alpha-mannosidase, use of the composition as a medicament, use as a medicament for the treatment of alpha-mannosidosis and a method of treating alpha-mannosidosis and/or alleviating the symptoms of alpha-mannosidosis.

The present invention comprises a method of concentrating a composition comprising a polypeptide of interest and the use of such concentrated composition for the treatment of diseases in mammals, in particular by subcutaneous injection.

The present invention comprises a method of concentrating a composition comprising a polypeptide of interest and the use of such a concentrated composition for the treatment of diseases in mammals, in particular by subcutaneous injection.

The present disclosure relates to compositions and methods useful for the production of recombinant glycoproteins in filamentous fungal cells, such as Trichoderma cells, wherein at least 90% (mol %), preferably at least 95% of the total neutral N-glycans of said produced recombinant glycoprotein are mammalian-like N-glycans. More specifically, the invention provides a filamentous fungal cell comprising i. one or more mutations that reduces or eliminates one or more endogenous protease activity compared to a parental filamentous fungal cell which does not have said mutation(s); ii.a polynucleotide encoding a heterologous catalytic subunit of oligosaccharyl transferase; iii. a recombinant polynucleotide for increasing 1, 2 mannosidase activity;and, iv. a recombinant polynucleotide encoding said heterologous glycoprotein.

Provided are a recombinant collagen, an expression method and use thereof. In the present disclosure, the recombinant collagen is expressed by a human embryonic kidney cell Expi293F at a high protein expression level, thereby realizing the expression of a large-molecular-weight recombinant collagen. Activity studies have showed that the expressed recombinant collagen has desirable activity in promoting cell migration, while the recombinant collagen III+I with a fusion tag shows high stability at 4 C. In addition, cell activity studies have shown that the recombinant type III collagen expressed by human cells has better activity in promoting the cell migration than that of a commercially available type III collagen. These results indicate that the human cell expression system is conducive to the expression of highly active and large-molecular-weight recombinant collagen.

Provided are compositions comprising vectors for the co-expression of a modified GlcNAc-1-Phosphotransferase gene and a lysosomal enzyme. The gene encoding the lysosomal enzyme is operably linked to a first promoter and the gene encoding the GlcNAc-1-Phosphotransferase is operably linked to a second promoter. Also provided herein are methods of treating a lysosomal storage disorder comprising administering to a subject the compositions of the disclosure.

The invention relates to means and methods for gene therapy of lysosomal storage disorders (LSDs), preferably a LSD with skeletal involvement, based on an ex vivo gene therapy approach comprising transduction of autologous hematopoietic stem and progenitor cells (HSPCs) with viral vectors for expressing enzymes that are deficient in the disorders. The final formulation is a suspension of transduced cells in culture medium for the administration to patients affected by the LSDs, preferably preceded by a conditioning regimen.