Abstract: A process for separating a protein from a sample containing a mixture of proteins is disclosed. The process comprises forming an aqueous combination comprising the sample and an ion exchange material at a pH which is less than the pI of a first fraction of the proteins in the aqueous dispersion but greater than the pI of a second fraction of the proteins in the aqueous dispersion. The aqueous dispersion may then be intensely mixed, causing a fraction of the proteins in the combination to bind to the ion exchange material, and, optionally, centrifuged, causing the combination to stratify into a concentrated solids fraction and a supernatant, the supernatant containing the fraction of the proteins that did not bind to the ion exchange material. This process, including the steps of intense mixing and centrifugation, requires no expensive, specialized instrumentation, can be quickly performed, and is carried out in non-denaturing conditions, allowing for the separated proteins to be further studied.

Abstract: The subject of the invention is separation on a solid support by multi-column sequential selective retention in order to separate a product of interest from a solution containing such a product of interest, by passing this solution over a fixed chromatographic resin bed comprising at least three zones, liquid flow means being placed between adjacent zones and between the last zone and the first zone, this process comprising several sequences, each sequence comprising at least one step chosen from an absorption step, a rinsing step and a desorption step, carried out either simultaneously or not, each following sequence is carried out by the displacement of the fronts in the zones towards the downstream end substantially of the same increment before the periodic displacement of the points of introduction and of withdrawal, characterized in that the process includes a sub-sequence without injection of the charge.

Abstract: The present invention relates to phosphopeptide isolation and enrichment, particularly to a highly selective and specific phosphopeptide enrichment method. The method mentioned in the invention is based on the strong interaction between zirconium phosphonate and phosphopeptide. Zirconium phosphonate is immobilized on the surface of matrix for phosphopeptide enrichment. Zirconium phosphonate could be immobilized on chip surface, it can also be immobilized on chromatographic matrix for isolation and purification of phosphopeptides. The method shows high specificity for phosphopeptides, it can be applied to purify and enrich low abundance phosphopeptide from biological samples.

Abstract: The present invention provides a purified multimeric complex form P. gingivalis. The complex comprises at least one domain from each of RgpA, Kgp and HagA, and has a molecular weight greater than about 300 kDa.

Abstract: A method of preparing a purified, virus inactivated and virus safe antibody preparation from a starting solution comprising antibodies and contaminants, the method comprising the steps of: (a) adjusting the pH of the starting solution to about 4.6 to about 4.95 in particular to about 4.8 to about 4.95 to produce an intermediate solution; (b) adding caprylate and/or heptanoate ions to the intermediate solution and maintaining the pH at about 4.6 to about 4.95 in particular pH at about 4.8 to about 4.

Abstract: The present invention relates to a process for purifying fibrinogen, which comprises one or more process steps in which one or more contaminating proteins are depleted by negative chromatography and/or negative adsorption using cation exchanger, hydrophobic gel and/or dye gel. In addition, the invention relates to the fibrinogen which is obtained by the process of the invention and which is notable for improved stability, and to the production and use of pharmaceutical preparations comprising this fibrinogen.

Abstract: A method for purifying an antibody by cation exchange chromatography is described in which a high pH wash step is used to remove of contaminants prior to eluting the desired antibody using an elution buffer with increased conductivity.

Abstract: The invention provided is a split-free nano-liter flow liquid chromatography system comprising at least a first and a second chromatographic column connected in series, wherein an autosampler is provided and programmed to inject samples and eluent solutions of increasing eluent concentration for gradually eluting analytes from the first column into the second column (without adversely affecting separation in the second column), said solutions segmented from the mobile phase of the liquid chromatography system by air plugs.

Abstract: The present application describes humanized anti-ErbB2 antibodies and methods for treating cancer with anti-ErbB2 antibodies, such as humanized anti-ErbB2 antibodies.

Abstract: This disclosure concerns compositions and methods for the treatment and inhibition of infectious disease, particularly bubonic and pneumonic plague. In certain embodiments, the disclosure concerns immunogenic proteins, for instance substantially monodisperse F1-V fusion proteins, that are useful for inducing protective immunity against Y. pestis.

Abstract: Processes and apparati are provided for separating molecules of interest from a mixture by depth filtration (DF). The DF of the invention is useful in the clarification and processing of various feedstreams for the removal of a molecule of interest. According to a preferred embodiment, a transgenic milk feedstream is stabilized and particulate matter such as fat, casein miscelles and bacteria are removed. An aseptic filtration step was also developed to remove any bacteria remaining in a clarified transgenic milk feedstream.

Abstract: A method for purifying a polypeptide by ion exchange chromatography is described which involves changing the conductivity and/or pH of buffers in order to resolve a polypeptide of interest from one or more contaminants.

Abstract: The invention is a method for the purification of mono-PEGylated erythropoietin using two cation exchange chromatography steps wherein the same type of cation exchange material is used in both cation exchange chromatography steps and a method for producing a mono-PEGylated erythropoietin in substantially homogeneous form.

Abstract: The present invention provides methods for purifying proteins. In particular, the methods employ a two-step non-affinity chromatography process without the use of an in-process tangential flow filtration step.

Abstract: The present invention relates to the purification of a Factor VII polypeptide from a bulk of a Factor VII polypeptide with respect to desirable glycoforms by fractionated elution from an anion-exchange material with an eluting buffer comprising a certain concentration of calcium ions. The present invention renders it possible to enrich a bulk of a Factor VII polypeptide with respect to desirable glycoforms.

Abstract: Substantially pure 2S canola protein is obtained substantially free from 7S and 12S canola protein by a procedure in which 2S canola protein is captured by binding to a cation-exchange medium while permitting other proteins and impurities to be washed away. The 2S canola protein then is removed from the cation-exchange medium by exposure of the cation-exchange medium to saline at a suitably high salt concentration.

Abstract: The present invention relates to a method for producing a mature von Willebrand Factor (VWF) from von Willebrand Factor pro-peptide comprising the steps: immobilizing VWF pro-peptide on an ion exchange resin, incubating the immobilized VWF pro-peptide with furin to obtain immobilized mature VWF, and isolating mature VWF from the ion exchange resin by elution.

Abstract: The present invention provides a method for recombinant production, recovery and purification of endostatin protein. This method may be employed for large scale recovery and purification of recombinantly-produced endostatin protein.

Abstract: A polymer carrier to which a prescribed zinc complex group is binding directly or through a spacer, having property to bond, under a certain condition, to an anionic substituent (a phosphate group, for example), showing low solubility to a solvent (preferably insolubility to a solvent) as a whole, and being capable of capturing, separating and purifying easily a substance having an anionic substituent (a phosphate group, for example).

Abstract: The present invention is directed generally to recombinant methods for making a desired pegylated protein and pooling of same. The method(s) yield a polypeptide product containing reduced levels of aggregate thereof pooled to provide the desired pegylated isoforms thereof.

Abstract: The present invention relates to a method for pre-fractionation of complex protein and/or peptide samples. The method comprises the following steps: a) loading the sample onto pre-packed media able to separate proteins/peptides according to their pl-value; b) eluting the media by centrifugal force under denaturing conditions with at least three buffers having different pH in a stepwise manner to obtain at least three fractions separated according to pl-value; and c) subjecting each fraction to further separation in a pH-range corresponding to the pl-values of said fractions. The invention also relates to a kit comprising at least one spin column or multiwell plate filled with chromatography medium able to separate proteins/peptides according to pl-value, at least three buffers, and one or more IPG strips having a narrow pH-range adapted to each of said buffers.

Abstract: The invention relates to the use of a factor H for producing a drug for treating Uremic Haemolytic Syndrome (UHS), to a method for purifying the factor H from a frozen fresh plasma and to a factor H concentrate obtainable by said method.

Abstract: The present invention generally relates to methods for separating albumin from samples comprising albumin and other proteins based on the pI of albumin. More particularly, the methods relate to forming an aqueous combination comprising the sample and an anion or a cation exchange material at various pHs, agitating the combination, and separating a fraction of unbound proteins from a concentrated solids fraction comprising the ion exchange material and bound proteins.

Abstract: The present invention relates to a protein found in natural rubber that can induce an allergic reaction in persons who have been sensitised to it. The invention provides for the process of isolating and purifying the protein and describes the characteristics of the protein, including its molecular weight, isoelectric point, amino acid sequence and allergenicity. The invention also describes the isolation and cloning a the DNA that encodes the protein. The production of the recombinant version of the protein using a protein expression vector is described.

Abstract: It is intended to provide a method whereby a protein can be separated and purified at a high accuracy by a convenient procedure. A sample containing the desired protein is brought into contact with an ion exchanger under first conditions at a high ionic strength and at pH value not in the vicinity of the isoelectric point of the desired protein. Next, the component adsorbed by the ion exchanger is eluted under second conditions at a lower ionic strength than in the first conditions and at a pH value closer to the isoelectric point of the desired protein than in the first conditions.

Abstract: The present invention relates to a method for obtaining recombinant granulocyte-colony stimulating factor (G-CSF), comprising at least one cation exchange chromatography and at least one hydrophobic interaction chromatography, wherein said two chromatographic steps are immediately consecutive in optional order. In particular, the present invention relates to a method for purifying G-CSF from a mixture of G-CSF and other proteins, comprising two cation exchange chromatography steps which are conducted before and after a hydrophobic interaction chromatography, respectively.

Abstract: Methods are provided for isolation of chromatin fractions of nucleoproteins containing histone H1, H2A, H2B, H3 and H4 proteins and/or histone H1, H2A, H2B, H3 and/or H4 proteins, from intact cells. The methods preserve original patterns of covalent modifications of the histone proteins.

Abstract: The present invention provides a process for manufacture of purified Nematode-extracted Anticoagulant Proteins (NAPs), wherein the NAP manufactured by the claimed process method is a NAP drug substance that can be formulated as a NAP drug product. The present invention provides NAP drug substances and NAP drug products manufactured by the process disclosed herein. In one embodiment, the present invention provides a process for manufacture of rNAPc2/proline drug substance and rNAPc2/proline drug product, and provides rNAPc2/proline drug substance manufactured by the process disclosed herein.

Abstract: Process for separating proteins fibrinogen, Factor XIII and biological glue from a solubilized plasma fraction and for preparing lyophilised concentrates of said proteins Summary The invention is related to a process for separating proteins fibrinogen, Factor XIII and biological glue from a solubilized plasma fraction and for preparing freeze-dried concentrates of said proteins comprising the steps of: chromatographic purification comprising the steps of loading an anion exchanger of weak base type with the said solubilized fraction, previously equilibrated with a buffer of a predetermined ionic strength of an alkaline pH, which allows to retain the biological glue, elution of the biological glue by increasing the ionic strength of the said buffer, and separation of FXIII from fibrinogen by addition to at least one part of the biological glue eluate of at least one chemical agent precipitating the FXIII, and recovery of the resulting purified fibrinogen containing supernatant solution, and diafiltration of

Abstract: This invention provides methods and compositions for producing high titer, substantially purified preparations of recombinant adeno-associated virus (AAV) that can be used as vectors for gene delivery. At the onset of vector production, AAV producer cells of this invention typically comprise one or more AAV packaging genes, an AAV vector comprising a heterologous (i.e. non-AAV) transgene of interest, and a helper virus such as an adenovirus. The AAV vector preparations produced are generally replication incompetent but are capable of mediating delivery of a transgene of interest (such as a therapeutic gene) to any of a wide variety of tissues and cells. The AAV vector preparations produced according to this invention are also substantially free of helper virus as well as helper viral and cellular proteins and other contaminants. The invention described herein provides methods of producing rAAV particles by culturing producer cells under conditions, such as temperature and pH, that promote release of virus.

Abstract: The present invention relates to a method for the purification of rFIX using anion exchange chromatography in the pseudo-affinity mode, wherein said method comprises a wash step with a wash buffer having a salt concentration of more than 200 mM. The purification according to the invention provides a method to enrich rFIX molecules which have been posttranslationally modified by sulfation and/or phosphorylation. The present invention further relates to purified rFIX compositions enriched in monosulfated and/or monophosphorylated rFIX molecules.

Abstract: The present invention describes a novel process for large scale purification of therapeutic grade quality of recombinant human G-CSF from microbial cells, wherein the protein is expressed as inclusion bodies. The process involves the novel use of Hydrophobic Interaction Chromatography (HIC) step to purify G-CSF eluted from a cation exchange column. A combination of these two chromatography steps provides good purity and yields which are essential for a production scale process. The host cell related contaminants like proteins, DNA and endotoxins are estimated to be within the specifications outlined by the drug regulatory authorities.

Abstract: The present invention relates to a method of separating antibodies from contaminants in a solution, which method comprises contacting the solution with a chromatography resin comprised of a support to which multi-modal ligands have been immobilised, wherein a multi-modal ligand comprises at least one cation-exchanging group and at least one aromatic or heteroaromatic ring system. In one embodiment, the ring-forming atoms of the aromatic or hereoaromatic entity are selected among C, S or O, and the cation exchanging group is a weak cation exchanger. The present method may be used as a single step procedure or as a polishing step following a capture on a Protein A column.

Abstract: The invention relates to methods of producing Protein A without contamination of the Protein A by animal products. The invention also relates to a vegetarian fermentation media in which Staphylococcus aureus is grown to produce a vegetarian Protein A. The invention further relates to a vegetarian Protein A and the use of a vegetarian Protein A in therapeutic and prophylactic methods.

Abstract: We describe a process for the monitoring of contaminant removal during the purification process of a pharmaceutical product produced by a host cell, wherein at least two different samples taken during the purification process of the pharmaceutical product are incubated with at least one protein biochip array and contaminants in the samples bound to the protein biochip array are subsequently detected. Preferably samples are taken before the first purification step and after each subsequent purification step.

Abstract: The present invention provides a method for purifying butyrylcholinesterase from various biological fluids. Biological fluids include, e.g., blood, blood fractions, plasma, and bioreactor broths, and other such mixtures containing butyrylcholinesterase. In one embodiment, the invention provides a method for the production of purified, virally inactivated butyrylcholinesterase by contacting a biological fluid containing butyrylcholinesterase with a cationic exchange chromatography material, with an affinity chromatography material, and treating the fluid with solvent detergent. The resulting purified butyrylcholinesterase can also be subjected to a pasteurization step, and formulated in a sodium chloride/sodium phosphate solution for storage or lyophilization.

Abstract: The expression vectors and methods using an E. Coli expression system for the large scale production of IL-29 are described. The vectors utilize the IL-29 coding sequence with specific changes in nucleotides in order to optimize codons and mRNA secondary structure for translation in E. coli. Also included are methods of producing, purifying and pegylating an IL-29 polypeptide.

Abstract: A separation material characterized by supporting, in its surface, a linear or crosslinked stimulus responsive polymer to which a substance A having specific affinity for a target substance and a low-molecular substance B having specific affinity for the substance A have been bonded. Such a separation material can increase the recovery of a target substance to be purified and do not require desalting and hapten sugar-eliminating operations and, in addition, have high productivity.

Abstract: The invention provides methods of purifying proteins, e.g., cytokines, from egg whites.

Abstract: 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.

Abstract: The present invention relates to methods and compositions for treatment of microbial infections and for the enhancement of resistance to infection. The invention comprises administration of an effective amount of bacterial lysate compositions for the treatment of pathological conditions of microbial infections. The present invention can also be used to enhance the immune system to prevent infections by the administration of an effective amount of the compositions.

Abstract: The present invention is an integral multilayered composite membrane having at least one ultrafiltration layer made by cocasting or sequentially casting a plurality of polymer solutions onto a support to form a multilayered liquid sheet and immersing the sheet into a liquid coagulation bath to effect phase separation and form a multilayered composite membrane having at least one ultrafiltration layer.

Abstract: Phosphorylated protein (i.e., phosphoprotein) affinity resins and methods for making and using the same are provided. The subject resins include a substrate bonded to aspartate-based tetradentate ligand/metal ion complexes, where the tetradentate ligand/metal ion complexes have high specificity for phosphorylated amino acids. The subject resins find use in a variety of different applications, including phosphoprotein enrichment applications. Also provided are kits and systems that include the subject resins.

Abstract: The present invention provides a process for manufacture of purified Nematode-extracted Anticoagulant Proteins (NAPs), wherein the NAP manufactured by the claimed process method is a NAP drug substance that can be formulated as a NAP drug product. The present invention provides NAP drug substances and NAP drug products manufactured by the process disclosed herein. In one embodiment, the present invention provides a process for manufacture of rNAPc2/proline drug substance and rNAPc2/proline drug product, and provides rNAPc2/proline drug substance manufactured by the process disclosed herein.

Abstract: The present invention relates to an ion exchange chromatography process for purifying a peptide from a mixture containing the peptide and related impurities, and to an industrial method including such ion exchange chromatography process.

Abstract: A novel protein and a process of producing the protein is provided. The protein is a glycoprotein having activity of suppressing the differentiation and/or maturation of adipocyte, having a molecular weight of about 45 kD under non-reducing conditions and about 28 kD and/or 23 kD under reducing conditions, and exhibiting affinity to heparin. A process of producing the protein comprising culturing human fibroblasts and purifying the culture broth by chromatography using an ion exchange column, affinity column, and reverse phase column. A cDNA encoding the protein and a process of producing the protein using the cDNA are also provided. The protein of the present invention is useful as a pharmaceutical composition for preventing or treating obesity or as an antigen for establishing immunological diagnosis, etc.

Abstract: The invention concerns a method for isolating milk proteins from milk or from whey comprising steps which consist in passing the milk or whey over a cation-ex-changing resin, eluting the fraction retained by an aqueous salted solution and desalting and sterilizing the eluate. The invention also concerns a milk protein fraction obtained by said method and its use for preparing pharmaceutical and food compositions.

Abstract: The present invention provides compositions and methods for selectively inhibiting the proliferation of stellate cells, which are important for the development of liver fibrosis upon liver injury. The invention describes conditioned media from immortalized hepatocytes as containing a death factor that induces apoptosis of activated liver stellate cells. This pro-apoptotic activity is shown to be associated with an 80 kDa protein, which is associated with a fetuin peptide sequence and an albumin peptide sequence.

Abstract: A process is disclosed for releasing proteins from cells and/or inactivating viruses. In the process, a host cell containing a protein of interest is contacted with a solution of an effective amount of a detergent.

Abstract: Compositions are described that are suitable for formulating TFPI. Solubilizers and stabilizers facilitate the preparation of pharmaceutically acceptable compositions of TFPI at various concentrations.