Recombinant vectors containing at least: a backbone comprising essential sequences for integration into a target cell genome; a nucleic acid encoding a CCR5 RNAi operatively linked to a a first expression control element that regulates expression of the nucleic acid encoding the RNAi of the CCR5; a nucleic acid encoding at least the extracellular domain of CD25 operatively linked to a second expression control element that regulates expression of the nucleic acid encoding at least the extracellular domain of CD25 are provided by this disclosure. In an alternative aspect, the vector also contains polynucleotides encoding TRIM5alpha and HIV TAR decoy sequences along with gene expression regulation elements such as promoters operatively linked to the polynucleotides. The vectors are combined with packaging plasmid and envelope plasmids and optionally conjugated to cell-specific targeting antibodies. Diagnostic and therapeutic methods for using the compositions are further provided herein.

Provided herein, inter alia, are methods, host cells, and vectors for producing 3-hydroxypropionate (3-HP). In some embodiments, the host cells include a recombinant polynucleotide encoding an oxaloacetate decarboxylase (OAADC) and a polynucleotide encoding a 3-hydroxypropionate dehydrogenase (3-HPDH). In some embodiments, the methods include culturing said host cell(s) in a culture medium comprising a substrate under conditions suitable for the recombinant host cell to convert the substrate to 3-HP. Expression of the OAADC and the 3-HPDH results in increased production of 3-HP, as compared to production by a host cell lacking expression of the OAADC and the 3-HPDH.

The disclosure provides a method of producing recombinant alkaline phosphatase comprising: (i) inoculating Chinese Hamster Ovary (CHO) cells expressing recombinant alkaline phosphatase in culture medium; (ii) culturing the CHO cells in the culture medium at a temperature of about 37° C.; (iii) adding a combination of nutrient supplements to the cell culture of (ii) at least one day after inoculation, the combination comprising (a) a first animal-derived component-free (ADCF) nutrient supplement comprising one or more amino acids, vitamins, salts, trace elements, poloxamer and glucose, wherein the first ADCF nutrient supplement does not comprise hypoxanthine, thymidine, insulin, L-glutamine, growth factors, peptides, proteins, hydrolysates, phenol red and 2-mercaptoethanol; and (b) a second ADCF nutrient supplement comprising one or more amino acids, wherein the second ADCF nutrient supplement lacks hypoxanthine, thymidine, insulin, L-glutamine, growth factors, peptides, proteins, hydrolysates, phenol red, 2-mercaptoethanol and poloxamer; (iv) decreasing the temperature of the cell culture of (iii) to about 30° C. about 80 hours to 120 hours after the inoculation; and (v) isolating the recombinant alkaline phosphatase from the cell culture of (iv) by at least one chromatography step.

The present disclosure is broadly concerned with the field of cancer immunotherapy. For example, the present invention generally relates to an immune cell comprising a genetically engineered antigen receptor that specifically binds to a target antigen and a genetic disruption agent that reduces or is capable of reducing the expression in the immune cell of two genes that weaken the function of the immune cell.

Provided is a method for producing a composition containing a polypeptide with suppressed coloring, the method comprising the steps of (a) culturing eukaryotic cells containing a nucleic acid encoding a polypeptide in a cell culture medium free of vitamin B12; and (b) collecting a composition containing the polypeptide from the culture.

The invention relates to a serum-free cell culture perfusion medium comprising the medium components subgrouped into at least three separate aqueous concentrated feeds and a diluent, wherein the resulting serum-free cell culture perfusion medium is pH-adjusting to neutral pH upon mixing. Also provided is a method of preparing said serum-free cell culture perfusion medium. The invention further relates to methods of culturing mammalian cells or producing a protein of interest in perfusion culture using said serum-free cell culture perfusion medium that achieve high productivity at a low cell specific perfusion rate. The invention further relates to the use of the new and improved serum-free cell culture perfusion medium to control osmolality in a perfusion cell culture, wherein increasing osmolality results in an increase in total productivity and/or cell specific productivity by suppressing cell growth during cell culture, e.g., during production phase of perfusion cell culture. Suppression of cell growth particularly reduces or eliminates the need for wasteful cell bleed.

The current invention reports the use of meta-tyrosine for increasing the specific productivity of a eukaryotic cell that produces/expresses a polypeptide. In the current method it is not necessary to perform a temperature-, osmolality- or pH shift or to add drugs like valproic acid or sodium butyrate to modulate the specific productivity of the cultivated cells. The method does not affect cell viability or product titer.

The present disclosure relates to genetically modified B cells, including memory cells, differentiated to plasmablasts or plasma cells useful for long term in vivo expression of a transgene, such as a specific antibody or other protein therapeutic. Also disclosed are methods of producing the cells and methods of treatment.

Antibodies and antigen binding fragments that specifically bind to human metapneumovirus (hMPV) F protein and neutralize hMPV are disclosed. Nucleic acids encoding these antibodies, vectors and host cells are also provided. The disclosed antibodies, antigen binding fragments, nucleic acids and vectors can be used, for example, to inhibit an hMPV infection or detect a hMPV infection.

Methods producing a recombinant protein of interest in a mammalian cell culture in media lacking IGF-1 are provided. Methods for producing mammalian cells capable of growing in media lacking IGF-1 are also provided.