Compositions, methods, articles of manufacture, and kits are provided for storage and delivery of proteins.

A composition comprising a protein and at least one single-stranded linear RNA oligonucleotide with an unfolded tertiary structure, which is characterized by: a) a relative content of each of the nucleotides A, C, U, and G, which is the number of each of the nucleotides per total nucleotide number in the oligonucleotide, which relative content is the same ±10% (number of nucleotides per total nucleotide number) as compared to the relative content of the respective nucleotide in a reference; and/or b) a relative content of at least 50% (number of nucleotides per total nucleotide number) of one type of nucleotide selected from the group consisting of nucleotides A, C, U, and G, which exhibits the highest relative content (number of each of the nucleotides per total nucleotide number) in a reference; wherein the reference consists of a native mRNA or a set of different RNA sequences that covers all RNA sequences allowed by the universal genetic code, each encoding an amino acid sequence of at least three amino acids within a p re-determined aggregation-prone target region of said protein, wherein the oligonucleotide has a length which is the same length as the reference, or longer.

A composition of renaturation buffer solution for dimeric proteins and method for using it. The renaturation buffer solution comprises buffer, oxido shuffling system, primary additives, chelating agent and polysorbate compound. The method for renaturation of dimeric proteins includes solubilization of target proteins using a solubilization buffer, solution dilution of the renatured target protein using a renaturation buffer containing polysorbate compound(s), concentrating of the target protein solution to a concentration approaching saturation. A method of applying renature denatured or misfolded proteins to become correctly folded and bioactive. A method of proper renaturation of recombinant proteins that have been expressed using translation vehicles (e.g., bacteria, insects, etc.) and proteins damaged by mechanical shearing, chemical stresses, and other stresses.

The present invention relates to methods for the enhancement and stabilisation of the proteolytic activity of proteases, methods for producing compositions comprising proteases with proteolytic activity that has been enhanced and stabilised, compositions comprising proteases with proteolytic activity that has been enhanced and stabilised obtained or obtainable by the aforementioned methods, use of such compositions in the manufacture of medicaments and cosmetics, the use of such compositions in the treatment of diseases and disorders including wounds, and in cosmetic applications, and associated kits.

The present disclosure pertains to the field peptide stapling and/or macrocyclization, where a structural motif is used to improve the properties of amino acid sequences (e.g. protease resistance, cellular penetration, biological activity). Also within the scope of the disclosure are methods for unstapling the S,S-tetrazine-containing amino acid sequence. The disclosure is also directed to methods for the reductive removal of thiocyanates from an amino acid sequence with cysteine to recycle back to the native amino acid sequence.

The present invention relates to a method of producing a collagen membrane that has particular mechanical properties. In particular, the present invention relates to a method A of producing a collagen membrane comprising the steps of (i) isolating a collagen-containing tissue and incubating same in an ethanol solution; (ii) incubating the collagen-containing tissue from step (i) in a first solution comprising an inorganic salt and an anionic surfactant in order to denature non-collagenous proteins contained therein; (iii) incubating the collagen-containing tissue produced in step (ii) in a second solution comprising an inorganic acid until the collagen in said material is denatured; and (iv) incubating the collagen-containing tissue produced in step (iii) in a third solution comprising an inorganic acid with simultaneous mechanical stimulation for sufficient time to enable the collagen bundles in said collagen-containing tissue to align; wherein the mechanical stimulation comprises applying tension cyclically to the collagen-containing tissue.

Disclosed herein is a protein purification system and methods of making such a system. Specifically, the invention relates to a method of immobilizing an N-terminal intein segment to a solid support, the method comprising: exposing an N-terminal intein segment to a cognate folding partner under conditions that promote association between the N-terminal intein and the cognate folding partner; immobilizing the N-terminal intein to a solid support; subjecting the N-terminal intein to conditions that disrupt association between the N-terminal intein and the cognate folding partner; and washing the solid support to remove non-bound material, thereby immobilizing an N-terminal intein segment to a solid support.

A method includes receiving one input feature and one output feature of importance of polymers used to stabilize a protein. A set of polymers from a library are identified based on the input feature and the output feature of importance that are applied to a machine learning model. Data for each polymer in the library includes features for each polymer and reagents for stabilizing the protein. Each polymer in the identified set is used to stabilize samples of the protein in well plates in a well plate array based on the reagents from the library data in the identified set. A score for each sample of the protein is assigned by comparing the measured output feature from the well plates corresponding to the identified set to the output feature of importance. The samples of the protein are identified having scores higher than a predefined threshold.

A method for crystallizing insulin or insulin analogs under alkaline conditions and purifying the insulin or insulin analog crystals by filtering through a filter and drying the insulin or insulin analog crystals captured on the filter to produce crystalline insulin or insulin analog crystal compositions is described. In particular aspects, the method may be used to crystalize insulin glargine.

The present invention provides methods of making α4β7 peptide monmer and dimer antagonists. Methods of the present invention include solid phase and solution phase methods, as well as synthesis via condensation of smaller peptide fragments. Methods of the present invention further include methods directed to the synthesis of peptides comprising one or more penicillamine residues.