Abstract: The present invention relates to a method of isolating an insoluble target protein from a suspension of intact or disrupted host cells. The invention also relates to insoluble target proteins which are obtainable by said method, in particular to silk proteins.

Abstract: The present invention relates to a method of isolating an insoluble target protein from a suspension of intact or disrupted host cells. The invention also relates to insoluble target proteins which are obtainable by said method, in particular to silk proteins.

Abstract: Processes are described herein for preparing medical devices and other articles having a low-fouling surface on a substrate comprising a polymeric surface. The polymeric surface material may possess a range of polymeric backbones and substituents while providing the articles with a highly efficient, biocompatible, and non-fouling surface. The processes involve treating the substrate to reduce the concentration of chemical species on the surface of or in the substrate without altering the bulk physical properties of the device or article, and thereafter forming a grafted polymer layer on the treated substrate surface.

Abstract: Methods for selective extraction and fractionation of algal lipids and algal products are disclosed. A method of selective removal of products from an algal biomass provides for single and multistep extraction processes which enable efficient separation of algal components. Among these components are neutral lipids synthesized by algae, which are extracted by the methods disclosed herein for the production of renewable fuels.

Abstract: The present disclosure includes a process for separating protein and an organic solvent from a defatted biomass comprising: a) contacting the defatted biomass with an aqueous solvent to: i) extract protein from the defatted biomass into the aqueous solvent and form an aqueous protein solution; and ii) reduce the concentration of the organic solvent in the defatted biomass and form an organic solvent fraction; b) separating the organic solvent fraction from the aqueous protein solution, wherein the organic solvent is present in the defatted biomass prior to the contacting step at a concentration of at least 100 ppm.

Abstract: A method is described for releasing a soluble or membrane associated intracellular protein of interest (POI) comprising the steps of: providing a cell comprising a soluble or membrane associated intracellular POI; contacting the cell with a membrane extracting composition; and causing the POI to be released from the cell under conditions sufficient for the specific release of the POI and in a soluble form.

Abstract: The present invention relates to a method for extracting organic molecules and/or organic minerals from a limestone material, including placing said limestone material in contact with a solution including at least one organic solvent, water and at least one acid, as well as to the organic molecules and/or organic minerals produced by the method. The products produced by the method can be used, for example, in therapeutic and non-therapeutic fields, in particular the agri-food, cosmetic and medical fields.

Abstract: A method for separating proteins from plant material, in particular, intact algal cells, using an amphipathic solvent set and a hydrophobic solvent set. Some embodiments include dewatering intact algal cells and then extracting proteins from the algal cells. The methods provide for single and multistep extraction processes which allow for efficient separation of algal proteins from a wet algal biomass. These proteins are high value products which can be used as renewable sources of food and food additives. Neutral lipids remaining in the algal biomass after extraction of proteins can be used to generate renewable fuels.

Abstract: Methods for selective extraction and fractionation of algal lipids and algal products are disclosed. A method of selective removal of products from an algal biomass provides for single and multistep extraction processes which enable efficient separation of algal components. Among these components are neutral lipids synthesized by algae, which are extracted by the methods disclosed herein for the production of renewable fuels.

Abstract: The present invention relates to a process of enriching one target compound from a liquid, which process comprises at least one step of isolation performed by differentially partitioning between two aqueous phases. In the present invention the phases are formed by adding a thermally responsive, self-associating (i.e. clouding) hydrophilic polymer, and if needed some additional salts, to an aqueous biotechnical solution (such as a fermentation sample or bioseparation process stream) under thermal and other conditions where the solution separates into a one polymer, two-phase system with one phase enriched in the polymer. The target compound is to be found in the phase not enriched in the polymer, while a significant though varying percentage of contaminants may differentially partition to the phase interface or the polymer enriched phase.

Abstract: The present disclosure includes a process for separating protein and an organic solvent from a defatted biomass comprising: a) contacting the defatted biomass with an aqueous solvent to: i) extract protein from the defatted biomass into the aqueous solvent and form an aqueous protein solution; and ii) reduce the concentration of the organic solvent in the defatted biomass and form an organic solvent fraction; b) separating the organic solvent fraction from the aqueous protein solution, wherein the organic solvent is present in the defatted biomass prior to the contacting step at a concentration of at least 100 ppm.

Abstract: A method for the production of a purified extract of natural allergens comprising the steps of a) extracting a natural source of allergens comprising allergenic proteins to form an extract, b) purifying of said extract to remove non-protein components to form a purified extract c) denaturating said purified extract to form a purified denaturated extract, said purified denaturated extract comprising proteins, wherein the most abundant (w/w) proteins, forming together at least 60% (w/w) of all proteins, are at least two proteins, and all proteins represent at least 60% (w/w) of the dry weight of the purified denaturated extract and a method for the production of a purified extract of natural allergens comprising the steps of a) hydrolysing a denaturated allergen to form an allergen hydrolysate, b) purifying said allergen hydrolysate to remove peptides with a molecular weight above 10,000 Da and below 1,000 Da in order to obtain a purified hydrolysate where 70%, more preferably 80% of the peptides are betwe

Abstract: We disclose methods for measuring vitamin D metabolite in plasma or serum samples. The methods comprise a step of adding to the plasma or serum samples a non-competitive displacement agent comprising 8-anilino-1-naphthalenesulfonic acid ammonium salt, 3-(acetonylbenzyl)-4-hydroxycoumarin and a water miscible solvent. The non-competitive displacement agent separates vitamin D metabolite from binding proteins in the sample, such that the displaced vitamin D metabolite is available for capture and detection in subsequent binding assays. Thus, our invention finds use in methods of separating and detecting vitamin D metabolites otherwise tightly bound to plasma or serum binding proteins.

Abstract: Blood collection, processing and transfer leads to the separation of discrete fractions by adding additional citrate (trisodium citrate) to bring the citrate concentration to 10%-15% w/v thereby leading to enhanced yield and purity of cryoprecipitate. The improved cryoprecipitate then yields concentrated clotting factors by an improved extraction process which uses polyvinyl pyrollidone to reduce the extraction of fibrinogen.

Abstract: Processes are provided for recovering and purifying refolded recombinant proteins produced in heterologous host cells, which includes the step of refolding the protein in a high pH buffer.

Abstract: A method is disclosed for extracting collagen from calcified tissue without a prior decalcification step. The method may be used with calcified tissues such as fish skin with scales, alligator skeletons, and crustacean exoskeletons. It does not require the use of EDTA. When fish skin is used that contains substantial quantities of scales, it is not necessary first to remove scales from the skin before conducting the extraction of collagen.

Abstract: A hydratable oxidized keratin composition comprising one or more metal ion species capable of absorbing water to form a hydrogel. The keratin material is useful as a soil amendment providing organic and inorganic nutrients. The keratin material is also useful as a nutrient source in the bioremediation of toxic contaminants soils and liquids.

Abstract: The present invention relates to use of avian antibodies and/or antigen binding fragments thereof, for the production of a drug for treatment and/or prevention of respiratory tract infection. The drug is administered through local application at the oral cavity and/or pharynx.

Abstract: The invention relates to a process of producing MAP-products from mussel feet. The process is characterised by extracting the mussel feet in a weakly acid aqueous solution containing, for instance, 1-10 percent by weight of a weak acid and 0.5-3 percent by weight of perchloric acid, whereafter the proteins in the aqueous solution are precipitated by adding inorganic or organic salts and separated from the system, subsequent to having extracted the solid substances.

Abstract: A hydratable, highly absorbent keratin solid fiber or powder capable of absorbing a large weight excess of water may be produced by partially oxidizing hair keratin disulfide bonds to sulfonic acid residues and reacting the sulfonic acid residues with a cation. The neutralized suspension can be filtered, washed, and dried, leaving keratin solid which can be shredded into fibers and further ground into powder. Addition of water to the solid produces a hydrogel. The powder or hydrogel may be useful as an absorbent material, as a therapeutic for skin, or as an excipient. Another use for the hydrogel is as a biocompatible viscoelastic filler for implant applications.

Abstract: Disclosed is a method of obtaining a highly soluble protein which method generally includes at least the step of contacting the protein with an amount of antioxidant suitable to raise the solubility of the protein, which method may also be utilized to raise the protein yield of the process. Antioxidants suitable for use in the present invention include substituted and unsubstituted quinones, anisoles, toluenes and tocopherols. Also disclosed is a highly soluble protein which includes a protein and added antioxidant. Further disclosed are food products made from a highly soluble protein. Finally, a method of processing food products is disclosed which at least includes the step of incorporating a highly soluble protein into the food product.

Abstract: A process for extraction of myelin basic protein from myelin containing tissue, such as central nervous system tissue, which process comprises the following steps:extraction of the myelin basic protein from myelin containing tissue with an organic solvent selected from the group consisting of chloroform and compounds having a polarity similar to that of chloroform;incubation of the organic phase in the presence of a lower aliphatic alcohol or propylene glycol;transfer of the myelin basic protein from the lower aliphatic alcohol/organic solvent mixture to an aqueous phase with the aid of hydrogen ions (protons); andrecovery of the purified myelin basic protein. The invention also relates to the product obtainable by the process.

Abstract: A composition is provided comprising about 0.1 to 15 mg/ml of a polypeptide in a buffer having a pH of about 7-12 comprising about 5-40% (v/v) of an alcoholic or polar aprotic solvent, about 0.2 to 3M of an alkaline earth, alkali metal, or ammonium salt, about 0.1 to 9M of a chaotropic agent, and about 0.01 to 15 .mu.M of a copper or manganese salt. The buffer is suitably used in a method for refolding improperly folded polypeptides.

Abstract: In the method described, a protein-containing substance is first taken up in an alkaline solvent to give a solution. Insoluble constituents of the substance are separated off, the solution is neutralized and desalinated, and then the proteins contained in the solution are concentrated. The solubilization or disintegration of the protein-containing substance is carried out at room temperature using homogenization equipment. The heat dissipated into the protein-containing substance during homogenization is simultaneously removed. The pH of the alkaline solvent during the decomposition is over 11.5 and/or decomposition is carried out in the presence of a detergent, in particular sodium dodecylsulfate (SDS).

Abstract: Disclosed is a method of obtaining a highly soluble protein which method generally includes at least the step of contacting the protein with an amount of antioxidant suitable to raise the solubility of the protein, which method may also be utilized to raise the protein yield of the process. Antioxidants suitable for use in the present invention include substituted and unsubstituted quinones, anisoles, toluenes and tocopherols. Also disclosed is a highly soluble protein which includes a protein and added antioxidant. Further disclosed are food products made from a highly soluble protein. Finally, a method of processing food products is disclosed which at least includes the step of incorporating a highly soluble protein into the food product.

Abstract: A composition is provided comprising about 0.1 to 15 mg/mL of a polypeptide in a buffer having a pH of about 7-12 comprising about 5-40% (v/v) of an alcoholic or polar aprotic solvent, about 0.2 to 3M of an alkaline earth, alkali metal, or ammonium salt, about 0.1 to 9M of a chaotropic agent, and about 0.01 to 15 .mu.M of a copper or manganese salt. The buffer is suitably used in a method for refolding improperly folded polypeptides.

Abstract: Recovery of the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof produced in the form of inclusion bodies from recombinant host cells is carried out by providing a washed recombinant host cell suspension, adding a detergent and subjecting the cells to mechanical disruption. A second detergent is added, followed by another mechanical disruption and centrifugation to a pellet. The pellet is resuspended in a buffer and subjected to another mechanical disruption. Inclusion bodies are washed, resuspended and then recovered. In addition, endotoxins and DNA are removed from inclusion bodies containing the 52/48 kDa tryptic fragment of vWF or peptide subfragments thereof by mechanically disrupting the inclusion bodies in an aqueous buffer containing a detergent. A washed pellet is formed from these mechanically disrupted inclusion bodies and dissolved in a denaturant.

Abstract: The present invention provides a method for purifying high yields of IgG immunoglobulins from an egg yolk by a single phase separation step using a nonionic detergent.

Abstract: The invention involves the supercritical or near-critical fluid disruption of microbial cells and extraction of intracellular components. First, a solvent that is a gas at ambient conditions and that has a critical temperature of between 0.degree. and 100.degree. C. is selected. This solvent is brought to near-critical pressure or higher and to near-critical temperature. The solvent then is combined with a slurry of cells to saturate the cells with the solvent under the prescribed conditions. Next, the pressure is released to cause a pressure drop which results in partial disruption of the cell membrane and release of solvent and other materials from the cell. Novel apparatus and associated methods are provided for carrying out the foregoing process continuously.

Abstract: The invention relates to fractionating a mixture of polyethylene glycol (PEG)-protein adducts having different degrees of PEG substitution by partitioning the PEG-protein adducts in a PEG-containing aqueous biphasic system according to the degree of PEG substitution. A new PEG-gm-CSF obtained by the process is useful in pharmaceutical compositions for use in therapeutic or diagnostic methods.

Abstract: In a process for the production of a soluble native protein, such as immunoglobulin or methionine-prochymosin, in which an insoluble form of the protein is produced by a host organism transformed with a vector including a gene coding for the protein, the insoluble form of the protein is reversibly denatured in an alkaline aqueous solution at a pH selected to promote dissociation of a group or groups of the protein involved in maintaining the conformation of the protein, and the protein is subsequently allowed to renature by reducing the pH of the solution below a pH effective to denature the protein to produce the soluble native form of the protein. The pH of the alkaline aqueous is suitably in the range 9.0 to 11.5.

Abstract: Disclosed herein is a novel glycoprotein derived from Ganoderma mycelia. The glycoprotein is free of human hemagglutination ability, and has immunosuppressive activities and a molecular weight of 16,000-18,000 as measured by SDS gel electrophoresis or 12,000-16,000 as measured by tricin-SDS gel electrophoresis. The glycoprotein is produced by culturing Ganoderma mycelia, extracting the resultant Ganoderma mycelia with an aqueous solvent, and then purifying the resultant extract. An immunosuppressive agent containing an effective amount of the glycoprotein is also disclosed.

Abstract: The production of recombinant chymosin is disclosed in which an insoluble form of chymosin precursor is produced by a bacterial host cell transformed by a vector including a coding sequence for said precursor. Solubilization of said insoluble form of chymosin precursor is accomplished using urea at a concentration of at least 7M or guanidine hydrochloride at a concentration of at least 6M prior to cleaving said precursor to form chymosin. Said solubilization preferably additionally involves the denaturation of said precursor in an alkaline aqueous solution, e.g., at a pH between a 9 and 11.

Abstract: An extraction and purification process for human gamma interferon (HuIFN-gamma) from recombinant host cells producing the same in insoluble form is described, which comprises suspending the cells in a buffer solution having pH from 6.0 to 8.5, optionally complemented with an osmoprotector; disrupting the cells in conditions which do not activate the endocellular proteases; separating and solubilizing the pellet containing the insoluble HuIFN-gamma by extraction with phosphate buffer having pH comprised between 6.5 and 8.5 and finally purifying the solubilized HuIFN-gamma by one or more chromatographic techniques. Such a process allows non-degraded HuIFN-gamma to be obtained, with a biological activity and chemical-physical properties unaffected vis-a-vis the native product and in high yield and purity.

Abstract: The present invention provides the process for treating the animal hairs by solubilization wherein the animal hairs can be treated in a short time without any complicated operations and special apparatuses, and the process for recovering the solubilized product of the animal hairs wherein said product can be recovered simply and efficiently from the solution thereof. The former is the process for treating animal hairs by solubilization which comprises solubilizing the animal hairs in a weak alkaline liquid medium in the presence of an oxidizing agent whose concentration is high. The latter is the process for recovering a solubilized product of animal hairs which comprises admixing a solution of said product with an organic acid or an aqueous solution thereof to precipitate said product.

Abstract: A process and apparatus are disclosed for the demineralization of ground bone particles or pieces of cancellous or cortical bone. The apparatus includes multiple solution reservoirs which supply solutions to be pumped into one or more columns filled with bone samples to be demineralized. Solvent outflowing from the column(s) can be monitored for pH, calcium ion concentration or conductivity as a basis for determining when demineralization is complete. During the delipidization phase of processing, lipid solute can be monitored by photometry, or a small assay of the eluent can be dropped into deionized water to see whether a precipitate develops. Flow through the apparatus, including the types and amounts of solutions to be flowed through the apparatus, can be manually or computer controlled.

Abstract: Methods are disclosed for separating hemoglobin from erythrocytes by contacting erythrocytes with a hypotonic buffer solution at a rate sufficient to render the release of hemoglobin from said erythrocytes without significant lysis. The hemoglobin is then separated from the erythrocytes. Methods are also disclosed for purifying hemoglobin solutions of DNA, endotoxins and phospholipids by contacting the hemoglobin solutions with an anion exchange medium.

Abstract: The present invention relates to processes for purifying annexines. The overall process includes the steps of preparing a homogenized cell preparation and adjusting the pH to about 8.0 to 10.0, adding at least one bivalent cation, adding a phospholipid, washing the insoluble cell residue to remove the soluble constituents, and extracting the annexines from the cell residue with a chelating agent. This process initially promotes the adsorption of the annexines onto the insoluble cell residue or membrane of the host organism. The adsorption step makes it possible to eliminate unwanted soluble matter from the homogenized material by simple washing. Desorption is accomplished by using a chelating agent.

Abstract: Recombinant structural and functional polymers are purified by lysing of the cellular host, separation of the solid materials, washing and extraction of contaminants using a detergent solution at elevated temperatures.

Abstract: A process is disclosed in which high purity protamine-DNA complexes are prepared by collecting nucleoprotamines specific developmental stages of a life form, specifically, amphibian, egg by low temperature processing. The process also includes the steps of sequential homogenization in a high concentration aqueous salt solution at a buffered low pH, followed by ultracentrifugation to remove insoluble matter. Either a crude mixture or pure isolate of the complexes may be produced. Pure isolates require aqueous chloroform extraction to isolate protein and to remove lipids. Lyophilization then removes chloroform and excess water. The isolate is then fractionated by single pass alumina chromatography. Dialysis against pure water removes salts. Repeated lyophilization removes excess water and concentrates single protamines and protamine-like proteins. The mixture may then be reconstituted with 5% weight/volume heterologous or homologous DNA, in order to shield from charge toxicity.

Abstract: An aqueous two-phase protein partitioning system is disclosed which employs polyvinylpyrrolidone as the upper phase and maltodextrin as the lower phase and provides a low-cost system for protein partitioning. The system can also be employed with the amion derivatives of chlorotriazine dyes, which bind in a noncovalent manner with the PVP and serve as a ligand for the proteins to be separated.

Abstract: A method for the preparation of a protein in a physiologically active or native form, which method includesproviding a source of protein in a solubilized form,and a cationic exchange medium;contacting the source of protein and cationic exchange medium; andrecovering the protein in a physiologically active form.

Abstract: A method for the preparation of proteins in biologically active form including providing a source of protein solubilized from inclusion bodies with a cationic surfactant; providing a weak denaturing agent; and contacting the solubilized protein with the weak denaturant in water in an amount sufficient to allow the protein to remain in a biologically active form.

Abstract: A method is provided for extracting human interleukin-4 (IL-4) from IL-4 expressing bacterial cells. The method comprises treating the bacterial cells with a deactivating agent, disrupting the cell membrane and recovering the IL-4.

Abstract: A liquid phase containing preS2+S antigen is separated into two phases after which the desired antigen is concentrated, washed and adsorbed and desorbed from a fumed silica to produce a product that is purified and concentrated in antigen:protein ratio and that is suitable for final purification.

Abstract: A method for the separation or purification of biopolymers comprises adsorbing the biopolymer on the surface of liquid oil droplets; and separating the adsorbed biopolymer from the oil droplets. The adsorbed biopolymer is separated from the oil droplets by mixing the droplets in an aqueous liquid, removing the lower aqueous phase and adding a fresh aqueous phase to the droplets, cooling the mixture to solidify and coalesce the oil and to cause it to release the adsorbed biopolymer to the fresh liquid, separating the fresh liquid and the biopolymer from the coalesced oil, separating the biopolymer from the fresh liquid.

Abstract: A method is provided for producing an effective adjuvant response or stimulating the immune response of a warm blooded animal which comprises administering to said warm blooded animal an effective amount of a composition comprising refined detoxified endotoxin in combination with a pharmaceutically acceptable carrier.

Abstract: A method for the solubilization and recovery of insoluble parasite protective antigenic factors associated with parasite material comprising solubilizing the antigenic factors with a non-ionic detergent and separating the solubilized material from undispersed residual material. The purified protective antigenic factors are useful as vaccines, particularly against malaria, and as diagnostic agents.

Abstract: A process for the separation from other cellular materials of heat agglomeration resistant water soluble nitrogen containing organic compounds such as plasmids, RNA's, mitochondrial DNA's, viral DNA's, chloroplast DNA's, other episomal DNA's and certain proteins. The process comprises heating cellular materials in a solution of lysing agent to lyse the desired cells and to agglomerate water soluble nitrogen containing compounds such as certain chromosomal DNA's which are not resistant to agglomeration; centrifuging the resulting product to remove water soluble agglomerated materials; separating the supernatant liquid and precipitating the water soluble agglomeration resistant organic compounds with a water soluble precipitant. The process also includes separating the agglomeration resistant water soluble nitrogen containing compounds from each other by means of exclusion chromotography.

Abstract: A method of purifying hydrolyzed protein compositions by contact with a substantially phase-incompatible fluid extractant is provided. A reduction in the concentration of chlorohydrins, measured as 3-monochloro-1,2-propanediol, in hydrolyzed protein compositions can be obrtained by contacting the hydrolyzed protein composition with such a phase-incompatible fluid extractant, e.g., ethyl acetate. The method allows removal of chlorohydrins without substantially affecting the organoleptic qualities of the hydrolyzed protein.