The present invention provides recombinant gram-negative host cells that do not degrade protease-sensitive recombinant proteins yet grow to high cell density, methods for the use of these host cells to produce high-quality recombinant proteins, including antibodies and antibody fragments, at high yield, as well as compositions and methods relating to periplasmic expression of recombinant proteins or polypeptides of interest in host cells.
The present invention relates to method of purifying charge-shielded proteins from a cell lysate or periplasmic releasate using hydrophobic interaction chromatography as a first chromatography steps. Also provided herein are compositions comprising charge-shielded proteins and methods of treatment using purified charge-shielded proteins.
B01D 15/32 - Bonded phase chromatography, e.g. with normal bonded phase, reversed phase or hydrophobic interaction
B01D 15/36 - Selective adsorption, e.g. chromatography characterised by the separation mechanism involving ionic interaction, e.g. ion-exchange, ion-pair, ion-suppression or ion-exclusion
B01D 15/42 - Selective adsorption, e.g. chromatography characterised by the development mode, e.g. by displacement or by elution
4.
METHODS OF PURIFYING CHARGE-SHIELDED FUSION PROTEINS
The present invention relates to method of purifying charge- shielded proteins from a cell lysate or periplasmic releasate using hydrophobic interaction chromatography as a first chromatography steps. Also provided herein are compositions comprising charge- shielded proteins and methods of treatment using purified charge-shielded proteins.
C07K 1/20 - Partition-, reverse-phase or hydrophobic interaction chromatography
A61K 47/64 - Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
Erwinia asparaginase. Methods herein produce asparaginase having high expression levels in the periplasm or the cytoplasm of the host cell having activity comparable to commercially available asparaginase preparations.
C12N 15/78 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Pseudomonas
C12N 15/67 - General methods for enhancing the expression
05 - Pharmaceutical, veterinary and sanitary products
42 - Scientific, technological and industrial services, research and design
Goods & Services
pharmaceutical preparations, namely, pharmaceuticals used for stimulating white blood cell production, the prevention and treatment of bone and skeletal diseases, and the treatment of cancer, cardiovascular, gastrointestinal, neurological, respiratory, ocular, dermatological, pulmonary, metabolic, infectious, and inflammatory conditions; pharmaceutical preparations for use in connection with protein therapies. Research and development in connection with protein therapies for use in pharmaceutical products; Laboratory services, and research and development in the field of pharmaceutical research, namely conducting expression of engineered proteins.
05 - Pharmaceutical, veterinary and sanitary products
42 - Scientific, technological and industrial services, research and design
Goods & Services
Pharmaceutical preparations, namely, pharmaceuticals used for stimulating white blood cell production, the prevention and treatment of bone and skeletal diseases, and the treatment of cancer, cardiovascular, gastrointestinal, neurological, respiratory, ocular, dermatological, pulmonary, metabolic, infectious, and inflammatory conditions; pharmaceutical preparations for use in connection with protein therapies. Research and development in connection with protein therapies for use in pharmaceutical products; Laboratory services, and research and development in the field of pharmaceutical research, namely conducting expression of engineered proteins.
8.
BACTERIAL LEADER SEQUENCES FOR PERIPLASMIC PROTEIN EXPRESSION
Provided herein are bacterial leader sequences for periplasmic expression of heterologous proteins, fusion proteins comprising bacterial leader sequences, and methods of expression of same.
C07H 21/04 - Compounds containing two or more mononucleotide units having separate phosphate or polyphosphate groups linked by saccharide radicals of nucleoside groups, e.g. nucleic acids with deoxyribosyl as saccharide radical
C07K 1/00 - General processes for the preparation of peptides
C07K 14/00 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof
Erwinia asparaginase. Methods herein produce asparaginase having high expression levels in the periplasm or the cytoplasm of the host cell having activity comparable to commercially available asparaginase preparations.
C12N 15/78 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Pseudomonas
C12N 15/67 - General methods for enhancing the expression
Provided herein are methods of production of recombinant Erwinia asparaginase. Methods herein produce asparaginase having high expression levels in the periplasm or the cytoplasm of the host cell having activity comparable to commercially available asparaginase preparations.
E. coliPseudomonadales Pseudomonadales host cells at high expression levels and having activity comparable to commercially available asparaginase preparations.
C12N 15/63 - Introduction of foreign genetic material using vectorsVectorsUse of hosts thereforRegulation of expression
C12N 15/74 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
C12N 15/78 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Pseudomonas
05 - Pharmaceutical, veterinary and sanitary products
Goods & Services
Pharmaceutical preparations, namely, pharmaceuticals used for the prevention and treatment of bone and skeletal diseases; pharmaceutical preparations; veterinary preparations.
05 - Pharmaceutical, veterinary and sanitary products
Goods & Services
(1) Pharmaceutical preparations, namely, anthrax vaccine; pharmaceutical and veterinary preparations for diseases and conditions associated with exposure to B. anthracis organisms or spores.
The present invention relates to the field of medicine, in particular, to the production of large amounts of a soluble recombinant polypeptide as part of a fusion protein comprising an N- terminal fusion partner linked to the polypeptide of interest.
C12N 15/00 - Mutation or genetic engineeringDNA or RNA concerning genetic engineering, vectors, e.g. plasmids, or their isolation, preparation or purificationUse of hosts therefor
C12N 15/67 - General methods for enhancing the expression
C12N 15/74 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora
17.
PROCESS FOR PURIFYING RECOMBINANT PLASMODIUM FALCIPARUM CIRCUMSPOROZOITE PROTEIN
The present invention relates to processes for purifying high-quality recombinant Plasmodium falciparum circumsporozoite protein at high yields. This process provides rCSP at high yields without the need for denaturing and refolding the protein. The present invention overcomes obstacles previously encountered in the field, including dimerization, aggregation, and N-terminal degradation of rCSP. The process provided by the invention is scalable, and can be applied to large fermentation batches. The invention also relates to stable liquid formulations of recombinant P. falciparum circumsporozoite protein, and processes for stably maintaining rCSP in a stable liquid formulation.
The present invention relates to the field of recombinant toxin protein production in bacterial hosts. In particular, the present invention relates to production processes for obtaining high levels of a recombinant CRM 197, Diphtheria Toxin, Pertussis Toxin, Tetanus Toxoid Fragment C, Cholera Toxin B, Cholera holotoxin, and Pseudomonas Exotoxin A, from a bacterial host.
C12N 15/78 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Pseudomonas
C12N 15/31 - Genes encoding microbial proteins, e.g. enterotoxins
C07K 14/00 - Peptides having more than 20 amino acidsGastrinsSomatostatinsMelanotropinsDerivatives thereof
The present invention relates to the field of recombinant protein production in bacterial hosts. In particular, the present invention relates to a production process for obtaining high levels of a recombinant CRM197 protein from a bacterial host.
C12N 15/78 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Pseudomonas
C12N 15/31 - Genes encoding microbial proteins, e.g. enterotoxins
C12P 21/02 - Preparation of peptides or proteins having a known sequence of two or more amino acids, e.g. glutathione
The present invention relates to the field of recombinant protein production in bacterial hosts. It further relates to expression of soluble, active recombinant protein by using secretion signals to direct the protein to the periplasmic space of a bacterial cell. In particular, the present invention relates to a production process for obtaining soluble hG-CSF protein from a bacterial host.
C12N 15/78 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Pseudomonas
The present invention relates to the field of recombinant protein production in bacterial hosts. It further relates to extraction of soluble, active recombinant protein from an insoluble fraction without the use of denaturation and without the need for a refolding step. In particular, the present invention relates to a production process for obtaining high levels a soluble recombinant Type 1 interferon protein from a bacterial host.
Provided herein are methods and compositions for expression of a nucleic acid construct comprising nucleic acids encoding a) a recombinant polypeptide, and b) a prototrophy-restoring enzyme in a host cell that is auxotrophic for at least one metabolite. In various embodiments, the host cell is auxotrophic for a nitrogenous base compound or an amino acid. The invention involves introducing an analogue into the growth media for the host cell such that the analogue is incorporated into the recombinant polypeptide or a nucleic acid coding sequence thereof. In various embodiments, the compositions and methods disclosed herein result in improved recombinant protein expression compared to expression of recombinant protein in an antibiotic selection system, or compared to expression of the recombinant protein in an expression system that lacks a metabolite analogue.
Methods of identifying and expressing an antibody variant are disclosed wherein the method comprises identifying a binding region in an antibody, fusing the binding region to a plurality of scaffolds of antibody constant regions to obtain antibody fragment variants, expressing the antibody fragment variants in organisms to form constructs and expressing the constructs carried by the organisms to form induced cultures, wherein the organisms are expressed in HTP mode.
C12N 15/62 - DNA sequences coding for fusion proteins
C12N 15/78 - Vectors or expression systems specially adapted for prokaryotic hosts other than E. coli, e.g. Lactobacillus, Micromonospora for Pseudomonas
G01N 33/50 - Chemical analysis of biological material, e.g. blood, urineTesting involving biospecific ligand binding methodsImmunological testing
24.
TRANSLATION INITIATION REGION SEQUENCES FOR THE OPTIMAL EXPRESSION OF HETEROLOGOUS PROTEINS
The present invention provides methods and compositions for producing heterologous protein with improved yield and/or quality. A library of randomized ribosomal binding site sequences is provided for the identification of a translation initiation region sequence optimal for expression of the heterologous protein. Also provided are novel ribosomal binding site sequences, and vectors and host cells having those sequences. The library of randomized sequences is useful for screening for improved expression of any protein of interest, including therapeutic proteins, hormones, a growth factors, extracellular receptors or ligands, proteases, kinases, blood proteins, chemokines, cytokines, antibodies and the like.
