384 results about "Protein aggregation" patented technology

Embodiments of the invention are directed to a method of estimating the physical stability of a protein formulation. A particular embodiment of the invention places the protein formulation under an agitational stress that causes the protein to aggregate at an accelerated rate. In one embodiment, the change in protein aggregation is monitored spectroscopically using Thioflavin-T. Embodiments of the invention then utilize a survival curve analysis to ascertain the relative physical stability of the different protein formulations under study. This method was used to develop novel surfactant-stabilized insulin formulations in a rapid, cost efficient manner, thus illustrating the utility of the inventive method to the discovery and development of pharmaceutical protein formulations.

The present invention provides improved methods for the reduction or removal of protein impurities from a complex cellular Streptococcus pneumoniae lysate or centrate comprising serotype 3 polysaccharides involving steps relating to post-lysis heating or pH adjustment. In certain methods, the lysate is heated for a time and at a temperature sufficient to denature proteins present in the lysate and cause their aggregation and precipitation. In one embodiment, the lysate is heated to at least 60° C. for at least 30 minutes to cause protein aggregation and precipitation, more particularly about 60° C. to about 70° C. for about 30 to about 50 minutes, and even more particularly about 65° C. for about 40 minutes. In other methods, the pH of the lysate or centrate is increased to at least 8.0 to improve filterability, more particularly about 8.0 to 8.4, and even more particularly about 8.2. In further methods, heating and pH adjustment steps are combined to cause the aggregation and precipitation of proteins as well as to improve filterability of the lysates or centrates. In other methods, the pH of the lysate or centrate is lowered to about 3.0 to about 5.0 to cause protein aggregation and precipitation. Such methods allow for the production of substantially purified serotype 3 polysaccharide-containing lysates or centrates.

The present invention generally relates to systems and methods for determining oxygen in a sample, or in a subject. In one aspect, the present invention is generally directed to an article exhibiting a determinable feature responsive to oxygen, such as oxygen-sensitive particles. The particles may exhibit a determinable change with a change in oxygen concentration, and such particles can accordingly be used to determine oxygen. For example, in one set of embodiments, the particles may be at least partially coated with a protein, such as hemoglobin, that is able to interact with oxygen. In some cases, the protein may aggregate under certain conditions (e.g., under relatively low oxygen concentrations), and such protein aggregation may be used, for example, to cause the particles to become aggregated, which can be determined in some way. In some cases, such aggregation may be irreversible; i.e., the degree of aggregation corresponds to the most extreme oxygen concentrations that the proteins were exposed to. Such articles may be used, for example, to determine oxygen within a sample, or within a subject, such as a human subject. For instance, the article may be formed as a skin patch, or administered to the skin of a subject, e.g., on the surface of the skin, within the dermis or epidermis, etc., to determine oxygen within the subject.

Disclosed are methods of preventing, treating, or diagnosing in a subject a disorder in protein folding or aggregation, or amyloid formation, deposition, accumulation, or persistence consisting of administering to said subject a pharmaceutically effective amount of inositol stereoisomers, enantiomers or derivatives thereof.

The present invention provides novel compositions and methods for removal of protein aggregates from a sample in a flow-through mode.

The present disclosure provides an effective method for the refolding of denatured proteins in solution so that properly folded, biologically active protein in solution is recovered in high yield. The refolding takes place at pressures between about 0.25 kbar to about 3.5 kbar, advantageously at about 1.5 kbar to about 3 kbar. Typically a chaotropic agent is present at a concentration which is not effective for denaturing protein at atmospheric pressure, and optionally, oxidation-reduction reagents can be incorporated in the refolding solution so that native intramolecular disulfide bonds can be formed where that is desired. The method is applicable to substantially all proteins, especially after solubilization and / or denaturation of insoluble protein aggregates, inclusion bodies, or abnormal oligomeric (soluble) aggregates.

A melt-processed protein composition formed from a protein, plasticizer, and an electrophilic reagent is provided. The electrophilic reagent, for instance, may be selected to undergo a nucleophilic addition reaction with free sulfhydryl and / or thiyl radicals to help minimize the formation of disulfide crosslinking bonds that could otherwise lead to protein aggregation during melt processing. To enhance the degree to which the electrophilic reagent can limit crosslinking, a plasticizer is also employed that helps to mediate the adsorption of the electrophilic reagent into the internal structure of the protein, where it can be more stably retained. Furthermore, the temperature and shear rate employed during melt blending may also be selected to be relatively low to help limit polypeptide dissociation, thereby minimizing the impact of aggregation and embrittlement.

The present inventors discovered that incorporating an Fc region and an antigen-binding domain whose antigen-binding activity varies depending on ion concentration into an antigen-binding molecule that binds to an aggregate-forming antigen produces an antigen-binding molecule that can preferentially clear protein aggregates in comparison to protein monomers from plasma. Use of antigen-binding molecules of the present invention allows various diseases stemming from target tissues to be treated target-tissue-specifically. Use of antigen-binding molecules of the present invention enables treatment of diseases caused by protein aggregates.

The present invention is based, at least in part on the discovery of therapeutic agents capable of preventing, inhibiting or modulating abnormal processing, misfolding or aggregation of protein. The therapeutic agents of the invention may prevent, inhibit or modulate the formation of inclusions. The therapeutic agents of the invention may also be capable of facilitating clearance and / or blocking the cellular toxicity of inclusions to treat or ameliorate disorders characterized by protein aggregation. Compounds which bind to structural motifs commonly found in protein aggregates, such as β-sheets, would represent strong candidates for such compounds and are therefore desirable.

Compositions useful for treating neurological disorders including neurodegenerative disorders associated with deleterious protein aggregation, aberrant protein folding, such as brain amylogenic diseases, and / or neurodegenerative autoimmune disorders are described. Methods of using said compositions also are described. In particular, methods to treat a neurodegenerative disorder such as Alzheimer's disease and a neurodegenerative autoimmune disorder such as Multiple Sclerosis are contemplated utilizing proteosomes and / or Glatiramer Acetate, wherein the GA is in a submicron emulsion or a nanoemulsion.

Disclosed are methods of preventing, treating, or diagnosing in a subject a disorder in protein folding or aggregation, or amyloid formation, deposition, accumulation, or persistence consisting of administering to said subject a pharmaceutically effective amount of inositol stereoisomers, enantiomers or derivatives thereof.

Disclosed is a stable pharmaceutically acceptable formulation containing a pharmaceutically acceptable amount of a protein. Also disclosed are methods for preparing such formulations and methods for inhibiting protein aggregate formation induced by physical stresses associated with processing, manufacture, shipping, and storing protein formulations, particularly freeze / thaw stress.

A novel treatment of Alzheimer's disease and other disorders involving protein misfolding or aggregation is provided by enhancing or sustaining an antibody response against predominantly directed against pathological protein aggregates or neo-epitopes present on pathogenic forms of said protein or protein complex. Furthermore, therapeutic methods are also described, wherein ex vivo stimulated antigen-selected peripheral blood lymphocytes are regrafted into the cognate donor.

It is intended to provide a food or a drink with a favorable flavor which has an acidic nature, contains protein and minerals, and has been stabilized in dispersion without showing any protein aggregation or the like even in the absence of a stabilizer. It is also intended to provide such an acidic protein food or drink which shows no protein aggregation even in the case of using not only minerals hardly soluble in aqueous media but also highly soluble minerals. By using an acid-soluble soybean protein, it is possible to provide an acidic mineral-containing food or drink which has a high stability without resort to a stabilizer as an essential component. Owing to the acidic nature, moreover, a product having a refreshing flavor, which is never achieved by neutral foods or drinks, can be prepared.

A concentrated, immunoglobulin composition for treating subjects vaccinated against or infected with a pathogenic microorganism, is made by (a) selecting a population of individuals previously vaccinated against antigens associated with the pathogenic microorganism; (b) identifying very high titre individuals by determining the level of specific antibodies immunoreactive with the pathogenic microorganism in the blood of the individuals; (c) combining blood from the very high titre individuals; and (d) purifying and / or concentrating the product of step (c). A concentrated immunoglobulin composition can include specific antibodies immunoreactive with a pathogenic microorganism, wherein the titre of specific antibodies is at least 5 times higher than the average titre of specific antibodies of a population of individuals previously vaccinated against antigens associated with the pathogenic microorganism. The composition has a relatively high protein concentration and a low percentage of protein aggregates. The pathogenic microorganism is preferably smallpox virus or vaccinia virus.

The invention discloses a method for monitoring beta amyloid protein aggregation process by aggregation-induced emission, belonging to the technical field of biomedical research and clinical detection. With the method, the high affinity between sulfydryl on cysteine and maleimide is utilized to specially combine the beta amyloid protein on an aggregation-induced emission probe. The beta amyloid protein aggregation process is monitored on the basis of aggregation-induced emission enhancement phenomenon. The method has the advantages of high sensitivity, high selectivity, good stability, low manufacture cost and the like and is easy to control. The A beta42 and A beta40 contents can be monitored in a micromole, even a nano-mole level, the detection speed of the method is improved by 15 times if being compared with that of the detection method which utilizes probes, such as ANS (8-aniline-1-naphthalene sulfonic acid) and the like. The beta amyloid protein aggregation process can be qualitatively and quantitatively monitored, the method has a good application prospect if being applied to the pathologic diagnosis of latent patients suffering from senile dementia.

The invention provides compositions, methods and uses comprising a scyllo-inositol compound that provide beneficial effects in the treatment of a disorder and / or disease including a disorder in protein folding and / or aggregation, and / or amyloid formation, deposition, accumulation, or persistence.

The present invention relates to processes for the removal of unwanted protein aggregates from antibody preparations. One process involves removal when the aggregate and antibody are very close in pI value (“Bind-Elute” process). Another process involves the removal when the aggregate and antibody are very close in net electric charge and retention time on ion exchange resin (“Bind-Washout” process). Using either process, at least 90% of the antibody is recovered and 70-90% of the aggregate is removed.

The invention relates to a novel method for treating a variety of diseases and disorders, including polyglutamine expansion diseases such as Huntington's disease, neurological degeneration, psychiatric disorders, and protein aggregation disorders and diseases, comprising administering to patients in need thereof a therapeutically effective amount of one or more deacetylase inhibitors. The invention is also directed to a transgenic fly useful as a model of polyglutamine expansion diseases, which may be used to test potential therapeutic agents.

An objective of the present invention is to provide methods for stabilizing proteins and methods for suppressing protein aggregation, which comprise the step of adding meglumine to the proteins. Another objective of the present invention is to provide agents for suppressing protein aggregation, which comprise meglumine. Still another objective of the present invention is to provide pharmaceutical compositions comprising antibody molecules stabilized by meglumine, methods for producing the pharmaceutical compositions, and kits comprising the pharmaceutical compositions.To achieve the objectives described above, the present inventors assessed the antibody-stabilizing effect of meglumine, an amino sugar. As a result, the inventors discovered that meglumine was useful as a stabilizer for antibody molecules and also as an excipient for freeze-dried preparations.

The subject invention concerns the reduction of protein aggregation in the neurons of a mammal through the use of a ketogenic treatment such as a ketogenic diet, a physical training regimen and / or administration of agents to increase fatty acid oxidation. Such ketogenic treatment can be useful in the reduction of certain aggregates including amyloid β peptide, polyglutamine containing huntintin protein, polyglutamine containing androgen receptor, polyglutamine containing atrophin-1, polyglutamine containing ataxins, α-synclein, prion protein, tau and superoxide dismutase 1 (SOD1).

Methods of identifying a compound that inhibit, enhance and / or misdirect assembly of a viral capsid include providing fluorescently-labeled capsid polypeptides labeled with a fluorescent dye that changes fluorescence at high concentration, wherein assembly of the capsid polypeptides into the viral capsid results in juxtaposition of at least one of the N-terminus and C-terminus of the capsid polypeptides. In vitro assembly reactions are performed utilizing the labeled capsid polypeptide in the presence of the at least one test compound, and an amount of fluorescence is detected at at least one time point following induction of assembly, wherein the induction of assembly is detected as a change in the fluorescence of the sample. It is then determined whether the change in fluorescence detected in the presence of the at least one test compound following induction of assembly is decreased, increased, or occurs at a fast rate when compared to the fluorescence detected in the absence of the at least one test compound, wherein such decrease is an indication that the at least one test compound interferes with assembly of the viral capsid.

The present invention relates to methods and assays for identifying agents capable of inhibiting the mutant huntingtin protein, inhibiting or reducing cell death, in particular cell death associated with polyglutamine-induced protein aggregation, which inhibition is useful in the prevention, amelioration and / or treatment of neurodegenerative diseases, and Huntington's disease more generally. In particular, the present invention provides methods and assays for identifying agents for use in the prevention and / or treatment of Huntingtons disease. The invention provides polypeptide and nucleic acid TARGETs and siRNA sequences based on these TARGETS.

The invention relates to self assembling beta-strand peptides for forming nanostructures, compositions containing the peptides, and methods of forming the peptides. The invention further relates to uses of these peptides in drug delivery and enhancement of drug solubility, biomolecule detection, and biocatalysis applications. The peptides of this invention are further useful in models of protein aggregation disease.

An object of the present invention is to provide a method for stabilizing a protein or a method for inhibiting aggregation of a protein, these methods including the step of adding meglumine to the protein. The present invention also provides a drug containing meglumine, which stabilizes protein, or a drug which inhibits protein aggregation. Furthermore, the object of the present invention is to provide a pharmaceutical composition containing an antibody molecule stabilized by meglumine, a method for preparing the pharmaceutical composition, and a kit containing the pharmaceutical composition. In order to solve the above-mentioned problems, the present inventors studied the antibody stability effect of a kind of amino sugar, meglumine. It was found that meglumine can be used as a stabilizer for antibody molecules and also as an excipient for lyophilized formulations.

Soluble variants of recombinant proteins produced in a prokaryotic host cell, where the high expression levels often cause the original proteins to aggregate into insoluble inclusion body aggregates. The variant polypeptides retain biological function while increasing protein solubility with comparable or higher recoverable levels of biologically active protein when expressed in a suitable expression host. Methods of identifying critical residues and substituting them are provided to produce the variants.

The invention discloses human papillomavirus (HPV) antigen formulations which prevent protein aggregation and show prolonged stability as aqueous solutions. These formulations comprise a salt (such as sodium chloride) and a non ionic surfactant (Polysorbate 80 such as Tween 803) in physiologically acceptable concentrations.

The invention discloses an application of a guanidino compound represented by formula I or a pharmaceutically acceptable salt of the guanidino compound in the preparation of medicines for treating nervous system degenerative diseases. R in the formula I is hydrogen, a carboxyl group or a -C1~C5 alkyl-carboxyl group. The guanidino compound has a substantial protection effect on hydrogen peroxide induced neonatal rat hippocampal nerve cell injures, has a substantial inhibition effect on in vitro beta-amyloid protein aggregation, and has potential clinic application values.

A method for making a milk protein concentrate can comprise heating a liquid milk composition at near its natural pH to a temperature that causes the whey protein in the milk to bind to the casein in the milk to form protein aggregates. After cooling, the liquid milk composition can be subjected to membrane filtration using a membrane filter having a large pore size. The protein aggregates are retained to yield a milk protein concentrate. The concentrate can be further treated to provide a concentrate having a lowered lactose content.