The present invention relates in part to nucleic acids encoding proteins, nucleic acids containing non-canonical nucleotides, therapeutics comprising nucleic acids, methods, kits, and devices for inducing cells to express proteins, methods, kits, and devices for transfecting, gene editing, and reprogramming cells, and cells, organisms, and therapeutics produced using these methods, kits, and devices. Methods for inducing cells to express proteins and for reprogramming and gene-editing cells using RNA are disclosed. Methods for producing cells from patient samples, cells produced using these methods, and therapeutics comprising cells produced using these methods are also disclosed.

Method for producing bioproducts from streams of organic material, comprising the following steps: (i) the physical-biological pre-treating of the organic stream with water and one or more mechanical steps; (ii) setting the pH value of the mixture between pH 5 and pH 9; iii) adding an inoculum of a natural anaerobic culture that releases organic compounds; iv) a first separation step which splits the mixture into a solid and a liquid fraction, after which an anaerobic fermentation processes the solid fraction, with formation of biogas and digestate; v) an aerobic treatment of the separated liquid fraction with biological conversion of the organic compounds to a protein-rich bioproduct; vi) a second separation step with separation of the formed protein-rich bioproduct; vii) recirculating the liquid phase; viii) drying the formed protein-rich bioproduct, such that in the end a dry, protein-rich bioproduct is obtained as well as the bioproducts biogas and digestate.

Method for producing bioproducts from streams of organic material, comprising the following steps: (i) the physical-biological pre-treating of the organic stream with water and one or more mechanical steps; (ii) setting the pH value of the mixture between pH 5 and pH 9; iii) adding an inoculum of a natural anaerobic culture that releases organic compounds; iv) a first separation step which splits the mixture into a solid and a liquid fraction, after which an anaerobic fermentation processes the solid fraction, with formation of biogas and digestate; v) an aerobic treatment of the separated liquid fraction with biological conversion of the organic compounds to a protein-rich bioproduct; vi) a second separation step with separation of the formed protein-rich bioproduct; vii) recirculating the liquid phase; viii) drying the formed protein-rich bioproduct, such that in the end a dry, protein-rich bioproduct is obtained as well as the bioproducts biogas and digestate.

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 present invention generally relates to a method of peptides' or polypeptides' modification by glycosylation. In particular, the invention relates to one pot synthesis of disaccharide glycan on to the acceptor substrate and thereby generating O- and/or S-glycosylated neo-glycopeptides including antimicrobial peptides by using multifunctional recombinant nucleotide dependent glycosyltransferase.

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.

One or more embodiments of the present invention are to provide a new means of improving the productivity of a target protein. The present inventors have identified a novel protein consisting of the amino acid sequence set forth in SEQ ID NO: 2 through an exhaustive analysis of the nucleotide sequence of chromosomal DNA of a yeast belonging to the genus Komagataella. Activation of a gene encoding the novel protein provides a cell having an improved productivity of a target protein.

One or more embodiments of the present invention are to provide a new means of improving the productivity of a target protein. The present inventors have identified a novel protein consisting of the amino acid sequence set forth in SEQ ID NO: 2 through an exhaustive analysis of the nucleotide sequence of chromosomal DNA of a yeast belonging to the genus Komagataella. Activation of a gene encoding the novel protein provides a cell having an improved productivity of a target protein.

1) A method for obtaining a concentrated protein-rich phase from residues of bioethanol production. 2.1) Previously, the separation of a protein-rich phase from whole stillage from bioethanol production has been achieved either by the addition of chemicals or by process steps that are complex in terms of equipment and/or energy. 2.2) Whole stillage from bioethanol production is fed to a solid-liquid separation, and the liquid phase (thin stillage) resulting from this is partially returned to the mashing process. This recirculation increases the raw protein content in the process. Part of the thin stillage is diluted and fed to a simple separation process without the addition of chemicals and temperature treatment, with a protein-rich phase being obtained. 2.3) A protein-rich phase is obtained from residues of bioethanol production.

1) A method for obtaining a concentrated protein-rich phase from residues of bioethanol production. 2.1) Previously, the separation of a protein-rich phase from whole stillage from bioethanol production has been achieved either by the addition of chemicals or by process steps that are complex in terms of equipment and/or energy. 2.2) Whole stillage from bioethanol production is fed to a solid-liquid separation, and the liquid phase (thin stillage) resulting from this is partially returned to the mashing process. This recirculation increases the raw protein content in the process. Part of the thin stillage is diluted and fed to a simple separation process without the addition of chemicals and temperature treatment, with a protein-rich phase being obtained. 2.3) A protein-rich phase is obtained from residues of bioethanol production.