Recombinant expression vectors are disclosed that include a control sequence for recombinant expression of proteins of interest; the control sequence combines a mCMV enhancer sequence with a rat EF-1alpha intron sequence. Some of the vectors are useful for tetracycline-inducible expression. Some of the vectors contain a 5′ PiggyBac ITR and a 3′ PiggyBac ITR to promote genomic integration into a host cell chromosome. A method of selecting a stable production cell line for manufacturing a protein of interest is also disclosed. Also disclosed are mammalian host cells comprising the inventive recombinant expression vectors and a method of producing a protein of interest, in vitro, involving the mammalian host cell.

A method of directly microbially converting a plant oil, an animal fat, free fatty acid, or a combination thereof to wax esters includes growing a yeast or bacterial strain in a medium comprising the plant oil, the animal fat, the free fatty acid, or combination thereof, under conditions suitable to produce the wax esters, wherein the yeast or bacterial strain is engineered to express a FAR gene encoding fatty acid alcohol reductase and a WS gene encoding a wax ester synthase, and optionally isolating the produced wax esters. Similar methods of directly microbially converting a plant oil, an animal fat, free fatty acid, or a combination thereof to omega-3 fatty acids by growing a microorganism in a medium comprising the plant oil, the animal fat, the free fatty acid, or combination thereof, under conditions suitable to produce omega-3 fatty acids are also described.

The present invention pertains to a cell culture medium comprising copper as a media supplement, which was shown to control recombinant protein glycosylation and methods of using thereof. The present invention further pertains to a method of controlling or manipulating glycosylation of a recombinant protein of interest in a large scale cell culture.

The present invention pertains to a cell culture medium comprising hypotaurine, GABA, and/or beta-alanine or the combination of choline and hypotaurine, GABA, and/or beta-alanine as media supplements which is shown to control viability, growth, and waste byproduct accumulation. The present invention further pertains to a method of producing a polypeptide of interest in a large scale cell culture containing hypotaurine, GABA, and/or beta-alanine or the combination of choline and hypotaurine, GABA, and/or beta-alanine.

The disclosure pertains to novel eukaryotic cell suitable for recombinant production of a product of interest, wherein the genome of the host cell is altered so that the effect of protein FAM60A is impaired in said cell, e.g. by reducing or eliminating functional expression of gene FAM60A thereby improving the stability characteristics. Furthermore, the present disclosure provides associated technologies wherein such host cells are used in recombinant production technologies.

The invention provides methods and materials for culturing mammalian cells and harvesting recombinant protein.

Methods of and system for growing and maintaining an optimized/ideal active yeast solution in the yeast tank and fermenter tank during the fermentation filling cycle are provided. A new yeast stage tank is used between the yeast tank and the fermenter tank allowing yeast to rapidly produce a huge amount of active young yeast cells for a fermenter during the filling period. A measurable and useful controlling factor, % DT/% Yeast by weight ratio (or “food” to yeast ratio), is used (e.g., % DT=glucose), which offers information on the health status of the yeast. The controlling factor is used to control the status of the yeast throughout the entire process.

A cell culture article is provided. The cell culture article includes a substrate of a polygalacturonic acid compound crosslinked with a divalent cation, the polygalacturonic acid compound being selected from at least one of: pectic acid; partially esterified pectic acid, partially amidated pectic acid and salts thereof. The substrate is digestible by digestion reagents into components including of galacturonic acid monomers and the divalent cation. Methods of harvesting cells from the cell culture article are also provided. The methods include performing a series of wash and/or centrifugation cycles to reduce components in the harvest solution.

Methods of improving recombinant protein titer and cell titer in cell culture using cell culture media having reduced impurities are provided, and well as cell culture media having reduced impurities that can used for the production of a recombinant protein and cells with improved titer. The cell culture media having reduced impurities comprises a HEPES buffer, and the reduced impurities are HEPES related impurities. In certain aspects, methods and media improve protein titer, cell growth, and/or viable cell density.

The present invention provides a device for producing a large number of uniform spheroids by an easy method. The spheroid-producing device (1) at least includes a first surface (11), a second surface (12), and a plurality of wall surfaces (13). The second surface (12) faces the first surface (11). The respective wall surfaces (13) constitute a plurality of holes penetrating through the first surface and the second surface. In addition, an equivalent diameter of inscribed circles of openings in the first surface (11) is greater than an equivalent diameter of inscribed circles of openings in the second surface (12).