Abstract: The present invention is directed to a protein purification construct having three tandem, coupled segments composed of a binding protein, an interconnecting linker and a variable fused polypeptide which incorporates the one or more copies of a product peptide. The binding protein is a mammalian or human carbonic anhydrase, or a modified version of the carbonic anhydrase. The protein purification construct may be employed in methods for expression of the product peptide in microbial and higher organism and for ligand immobilized affinity purification of the product peptide.

Abstract: The present invention aims at providing a method for preparing dermonecrotic toxin, an improved toxoid of dermonecrotic toxin produced by Bordetelia and a toxoid mixture comprising said improved toxoid of dermonecrotic toxin produced by Bordetella and a toxoid of dermonecrotic toxin produced by Pasteurella. There is provided a method for partially purifying dermonecrotic toxin which comprises bringing a dermonecrotic toxin-containing solution into contact with a chromatographic gel sulfated by direct sulfation or a chromatographic gel to which a sulfated molecule is covalently bonded to thereby make the dermonecrotic toxin adsorbed on the gel, and eluting the adsorbed dermonecrotic toxin from the gel.

Abstract: Liquid and lyophilized reagents for determining prothrombin time and/or fibrinogen levels in a plasma sample are disclosed. The reagents preferably are based on recombinant rabbit tissue factor. Also disclosed is a purified preparation of recombinant rabbit tissue factor having unique properties, and a novel method for making PT reagents.

Abstract: The present invention provides an enzyme L-fucokinase isolated from porcine kidney and in homogeneously purified form. The enzyme has a native molecular weight of 440 kDa based on gel filtration, a subunit molecular weight of about 110 kDa based on SDS PAGE, an optimal pH of about 8.0, and wherein said enzyme catalyzes the phosphorylation of L-fucose and does not phosphorylate D-glucose, D-galactose and D-mannose.

Abstract: To provide a method of efficiently separate protective components of Bordetella pertussis. On the basis of differences in adsorbability to calcium phosphate gel formed by adding calcium ions to a Bordetella pertussis culture in the presence of excess phosphate ions, protective components of Bordetella pertussis are separated from the Bordetella pertussis culture. Traditionally, protective components of Bordetella pertussis have been separated using different purification methods for the respective components. According to the present invention, the use of the same means of purification for all subject components makes it possible to purify each component with high efficiency and high recovery rate, an aspect very advantageous for industrial production. It is also possible to efficiently produce an improved purified pertussis component vaccine comprising an effective combination of pertussis filamentous hemagglutinin (FHA), pertactin (PRN, 69K-OMP), pertussis fimbriae (FIM) and pertussis toxin (PT).

Abstract: An adsorbent for a serum amyloid protein, which comprises a water-insoluble carrier and either a compound selected from the group consisting of a compound having an anionic functional group, a polyanionic compound having more than one anionic functional groups, aniline and an aniline derivative, said compound being immobilized onto said carrier, or a group of the formula: --NR.sup.1 R.sup.2, wherein R.sup.1 is hydrogen atom, methyl group or ethyl group, and R.sup.2 is an atomic group satisfying that the value of log P, in which P is a distribution coefficient in a water-octanol system, of a compound of the formula: R.sup.2 H is from 0 to 3.2, said group being bonded to said carrier, and a method for removing a serum amyloid protein from body fluid through a removing apparatus using the above adsorbent. According to the present invention, SA proteins, particularly SAA proteins and SAP proteins can be selectively removed from body fluid without decreasing HDL largely.

Abstract: The invention deals with a method for crystallizing in increased yields a polypeptide or a protein obtained from a protein solution comprising more than one protein, comprising:(a) treating the protein solution with a solid adsorption material; and(b) crystallizing the polypeptide or the protein after said solid adsorption material has been removed; or(c) crystallizing the polypeptide or the protein without removing said adsorption material.

Abstract: Mature alveolar surfactant protein B and a process for producing the same is disclosed. The process includes the use of a fusion protein of SP-B having a propeptide only at its amino terminus. A propeptide at its carboxyl terminus is not necessary, and thus not included in the fusion protein, to produce the mature SP-B of this process. Cleavage of the amino-terminus propeptide results in the production of mature SP-B.

Abstract: The present invention relates to cartilage extracts and to a method of producing the same. Shark cartilage extracts having anti-angiogenic, direct anti-tumor proliferating, anti-inflammatory and anti-collagenolytic activities have been obtained by an improved process. The process comprises the steps of obtaining a homogenate of cartilage in an aqueous solution, this homogenate being centrifuged and further fractionated to obtain a total extract having molecules of a molecular weight comprised between 0 to 500 KDa. The composition of the liquid extract has then been investigated by different ways. Further fractionation of this extract led to the preliminary characterization of some of its active components. Due to the multiplicity of biological activities of the total liquid extract, it can be used for treating numerous diseases or conditions such as those having components selected from the group consisting of tumor proliferation, angiogenesis, inflammation and collagenolysis.

Abstract: The invention relates to a pharmaceutical composition for the treatment of patients with blood coagulation diseases which are caused by coagulation factor deficiency and/or inhibitors of coagulation factors, whereby the composition as a FEIB-activity and is characterized in that it has Factor VIIa and at least one further active ingredient and the activity of at least 10 Factor VIIa units per unit FEIBA. Additionally, the invention comprises a method for the production of the pharmaceutical preparation and its use.

Abstract: The present invention relates to a process for purifying recombinant coagulation factor VIII by loading an aqueous solution containing factor VIII onto a hydrophobic interaction chromatography (HIC) gel, wherein at least one surfactant is present in the aqueous solution and/or a buffer solution used to equilibrate the gel prior to loading the aqueous solution onto the gel. The presence of a surfactant when loading the solution containing factor VIII onto the HIC gel, makes it possible to efficiently separate the intact and active factor VIII molecules from molecules with structural deviations. With the present invention it is further possible to reduce the content of DNA and/or CHO cell contaminants considerably and increase the activity of the factor VIII product to a hitherto unknown extent.

Abstract: The invention provides processes for producing Alpha-Interferon (IFN-.alpha.) free from possible mouse and/or virus contamination. The present invention further provides homogeneous IFN-.alpha. free from mouse and/or virus contamination and its use in antitumor and/or antiviral treatment.

Abstract: The instant invention relate to a new method for the purification of proteins using copper chelate-affinity chromatography, wherein the impure or pre-purified protein is adsorbed on immobilized copper(II) ions, optionally washed with buffer and deionized water, washed with a solution of a Lewis-base, and finally eluted with deionized water.

Abstract: The present invention provides a method for purifying follicle stimulating hormone (FSH) from biological samples, for example, from human pituitary glands or human postmenopausal urine, wherein the FSH is contaminated with other proteins, by use of dye-ligand affinity chromatography (DAC). This process may be used to generate affinity pure FSH suitable for therapeutic applications.

Abstract: The present invention relates to shark cartilage extracts and to a method of producing the same, these extracts having anti-angiogenic properties (reduction of the area of blood vessels observed in vivo on experimentally induced tumors), tumor regressive activity in vivo as well as demonstrating a direct inhibitory effect on tumor cell lines. This process does not involve any denaturing solvent or product and does not involve the use of any enzymes. It consists of obtaining a blend of whole cartilage in an aqueous solution of neutral pH, preferably pure water, this blend being centrifuged and the pellet and supernatant kept for further processing. The pellet is lyophilized and tested for anti-tumor and anti-angiogenic activities in vivo and in vitro, with or without supernatant. The supernatant has been shown to have anti-angiogenic and tumor regressive activities in vivo. The composition of the supernatant has then been investigated by different ways.

Abstract: The present invention is directed to a process for purifying alpha-1-PI. The process comprises providing an impure protein fraction which comprises alpha-1-PI. The impure protein fraction is suspended in an aqueous solution at pH 6. Insoluble proteins are recovered and resuspended in aqueous solution at pH 8.5. PEG is added to precipitate .alpha.-2 proteins. To the PEG supernatant precipitation, which comprises alpha-1-PI, is added ZnCl.sub.2 to precipitate crude alpha-1-PI. The crude alpha-1-PI is resolubilized and applied to an anion-exchange medium. A fraction comprising alpha-1-PI is recovered from the anion-exchange medium. Alpha-1-PI purified by the process has a specific activity about 1.0 units/OD.sub.280.

Abstract: The invention provides a process for purifying recombinant human serum albumin (rHSA) by heating a culture medium containing rHSA and the rHSA-producing host cello, feeding said heated solution upwardly into a fluidized bed in which adsorbent particles are suspended to effect contacting with the adsorbent particles and then recovering the adsorbed fraction containing the rHSA, and a composition comprising rHSA which shows a A35D/A280 ratio of below 0.015, when formulated into a 25% solution of said albumin.

Abstract: The subject invention provides a method of producing enzymatically active CPB which comprises treating a recombinant cell containing DNA encoding ProCPB, so that the DNA directs expression of the ProCPB, recovering from the cell the ProCPB so expressed, treating the recovered ProCPB under conditions permitting folding of the ProCPB, subjecting the folded ProCPB to enzymatic cleavage to produce enzymatically active CPB and purifying the enzymatically active CPB.

Abstract: A new use of hydrophobic zeolites, viz. for removing preservatives from (poly)peptide solutions, e.g. solutions of pharmaceutical preparations. The use is of particular interest in connection with protein solutions, especially insulin solutions. The invention further relates to an injection syringe for such solutions, in which syringe a zeolite of the type at issue is placed for contact with the solution to remove preservatives therefrom.

Abstract: A process for the removal of endotoxin from a biological product such as proteins for therapeutic use, blood plasma fractions, and albumin solutions that contains endotoxins, which comprises contacting said biological product with a cross-linked hydrophilic matrix comprising a copolymer of allyl dextran and N,N'-methylene bisacrylamide under conditions effective to bind endotoxins in said biological product to said matrix, wherein the total amount of endotoxin in said biological product does not exceed the absorptive capacity of said cross-linked hydrophilic matrix, and recovering purified biological product from which endotoxin has been removed as a result of said binding.

Abstract: The invention relates to a method for purification of a mixture of hydroxamate derivatized protein/proteins and native protein which is characterized by treating the mixture with immobilized metal and thereby adsorbing the hydroxamate derivatized protein/proteins on the immobilized-metal and recovering the native protein. The protein could be IGF-L. It also relates to a process for the production of a native protein which is characterized by expression of the protein as a fusion protein, cleavage of the fusion protein by hydroxylanine, separation of native protein from hydroxamate derivatized protein by adsorbing the hydroxamate derivatized protein on immobilized metal and directly recovering the native protein. The use of immobilized metal affinity chromatography for separation of native protein from hydroxamate derivatized protein is also claimed.

Abstract: Disclosed is a method for measuring or detecting inorganic phosphate ion in a sample. In the method, a sample is reacted with a phosphate-binding protein containing (i) an inorganic phosphate ion binding site and (ii) a detectable label that produces a signal whose amplitude changes detectably upon the binding of inorganic phosphate ion to the binding site, under conditions effective to allow binding of phosphate in the sample to the binding site. From a change in signal, the level of inorganic phosphate ion is measured or detected. Also disclosed are phosphate binding proteins for use in the method.

Abstract: The present invention relates to a pituitary adenylate cyclase activating polypeptide (PACAP) receptor protein or a salt thereof which is capable of binding PACAP and a method for preparing said receptor protein or salt thereof.

Abstract: A fimbrial agglutinogen preparation is prepared from a bordetella strain, particularly a B. pertussis strain, by a multiple step procedure involving extraction of the fimbrial agglutinogens from cell paste and concentrating and purifying the extracted material. The fimbrial agglutinogen preparation may be used to prepare acellular pertussis vaccines with other pertussis antigens, including pertussis toxin or toxoid thereof, the 69 kDa protein and filamentous hemagglutinin and other Bordetella antigens.

Abstract: Recombinant rabbit tissue factor is cloned and produced in a bacterial host. This protein, which is relatively insoluble and has several disulfide bonds, requires special modifications in order to express and be purified at commercial levels. By expressing the tissue factor as a fusion protein with a bacterial enzyme, thioredoxin, solubility is increased. Use of a thioredoxin reductase deficient host aids in proper tertiary structure for biological activity.

Abstract: A method for preparing an aqueous protein composition from human blood plasma, wherein the plasma undergoes a series of treatments by heating and precipitating agent to give albumin solutions containing the precipitating agent, and said precipitating agent is separated from the resulting solutions to give a crude aqueous albumin solution which is subjected to at least one liquid phase chromatography step to retain at least part of the secondary proteins other than albumin. The chromatography comprises affinity chromatography on a particulate support consisting of neutral particles loaded with at least one compound comprising sulphate groups, whereby, after the albumin solution has been fed through, the particulate affinity chromatography support is eluted by feeding through an aqueous saline solution, preferably by increasing the ionic strength, and the desired protein composition is collected by elution.

Abstract: The present invention, using a readily available sulfated chitin as an adsorbent, can permit platelet factor-4 to be recovered through specific adsorption from a solution containing the same factor, in by far increased yields as compared with the conventional process utilizing a heparin-immobilized affinity column, and provides the process for isolating, through purification platelet factor-4 which is suited for a commercial-scale, mass production process, wherein there can be offered the advantages of utilization of more readily available sulfated chitin, simplified procedure and improved production yields for the objective substance.

Abstract: Porous zirconia particles of specific gravity of 2.5-3.5 g/cm.sup.3 and mean particle sizes of 30-400 .mu.m can be synthesized using oil emulsion methods from colloids and used for protein adsorption in stable expanded beds. Expanded beds of less than 1.0 settled bed height to diameter (approximately 10 ml bed volume) are stable at linear fluid velocities of at least about 100 cm/hour.

Abstract: An adsorbent for removing ketoamine-containing protein, which comprises carrying a compound having a terminal functional group of the formula: ##STR1## wherein each of R.sup.1 and R.sup.2 is an organic group and X is 0 or 1, on a porous water-insoluble carrier, a process for removing ketoamine-containing protein by employing the above adsorbent, a process for preventing or treating for diabetic complication by removing ketoamine-containing protein with the above adsorbent and an adsorber for removing ketoamine-containing protein.

Abstract: The invention relates to a polysaccharide toxin from group B .beta.-hemolytic Streptococcus (GBS) having improved purity. The improved purity of the toxin, 95%, is achieved by the method of purification wherein a bacterial fermentation stock is subjected to chromatography with hydrophobic interaction chromatography (HIC) resin, extraction with a phenol/saline solution followed by ion exchange chromatography. The purified GBS toxin has a molecular weight of about 300 kD and a relative carbohydrate ratio of rhamnose:mannose:galactose:glucose:N-acetyl glucosamine:N-acetyl galactosamine of about 0:1:3:1:1:1, respectively. The GBS toxin is useful for treatment of tumors and other medical conditions.

Abstract: A solid sorbent material comprising cellulose which has been modified by hydrolysis with a cellulase enzyme for a duration sufficient to increase the protein adsorption capacity of the solid sorbent material and methods for preparing the sorbent material. Methods for purifying a protein include passing a liquid medium containing the protein over the solid sorbent material are also included.

Abstract: This application describes a high throughput assay for screening for compounds capable of binding to a fusion protein which consists of a target protein and an FK506-binding protein. The method for preparing the DNA encoding for the fusion protein and for expressing that DNA is also described in the application. The invention also discloses the recombinant DNA and protein sequences for several fusion proteins.

Abstract: A process is disclosed for preparing human inter-alpha-trypsin inhibitor (ITI) from human plasma which comprises two steps of anion exchange chromatography followed by a step of affinity chromatography on immobilized heparin. The process also includes a viral inactivation treatment. The quality of the ITI concentrate obtained is suitable for therapeutic use.

Abstract: The present invention provides a method for the purification and characterization of MHC-peptide complexes useful in ameliorating immunological disorders, such as, for example, autoimmune diseases, allergic responses and transplant responses. The method disclosed is a one-step method based on the use of metal chelate affinity chromatography to separate the MHC-peptide complexes of interest from both uncomplexed MHC molecules and other endogenous MHC-peptide bound complexes.

Abstract: Methods of isolating components of interest from a milk sample are described. The methods include a step wherein the solubility of at least a portion of the total milk protein is stabilized in such a manner as to allow isolation of the component of interest without significant loss in yield. Kits for stabilizing the solubility of at least a portion of the total milk protein of the milk sample containing the component of interest also are described.

Abstract: Methods are disclosed for the production and purification of hydrophobic fusion proteins production and purification of said hydrophobic polypeptides, proteins or peptides. Homogeneous monomeric .beta.-amyloid peptide and tests for screening amyloid toxicity-inhibiting drugs using this monomeric .beta.-amyloid peptide relate to these fusion proteins.

Abstract: The present invention is directed to novel purified and isolated mite allergens possessing mite allergen activity with a molecular weight of about 94,000, about 40,000, about 16,000 or about 14,000 as determined by SDS-PAGE, which can be isolated from extracts of mite, and to a process for producing such mite allergens. The purified and isolated mites allergens of the present invention are useful as a pharmaceutical and a diagnostic composition for mite allergic diseases.

Abstract: A vaccine effective against infection caused by group B Nesseria meningitidis microorganism is provided which comprises a purified protein antigenic complex weighing from 65 to 95 kDa, vesicles, and capsular polysaccharides. This vaccine is extracted from the cell membranes of the live microorganisms using detergent and enzyme. The method of producing an antimeningococcic hyperimmune gammaglobulin and the gammaglobulin produced by the method was provided. The gammaglobulin is obtained from vaccinees vaccinated with the vaccine.

Abstract: A method of preparing polyclonal immunoglobulin Fab fragments comprising cleaving immunoglobulin molecules, characterised in that the said cleavage is carried out on immunoglobulins in blood, serum or plasma in a closed, sterile environment.

Abstract: The present invention relates to a method and to a device for separating plasma from whole blood. The method and device utilize a permeable non-glass fiber matrix containing a polyol which is capable of clumping red blood cells. The matrix, in the absence of such a polyol, would otherwise be porous to red blood cells. The polyol-containing matrix has a first surface and a second surface such that a whole blood sample which is applied to the first surface flows directionally toward the second surface. Plasma separated from whole blood becomes available at the second surface of the matrix and can be tested for the presence of a particular analyte, such as glucose or fructosamine, as provided by multi-layer test devices of the present invention.

Abstract: A pseudobioaffinity chromatography adsorbent adapted for use in selectively adsorbing immunoglobulins. In one embodiment, the adsorbent includes: (a) a solid support material; and (b) a ligand immobilized on the surface of the solid support material, said ligand being a compound of the formula ##STR1## wherein Y.sub.1 is selected from the group of S, SCH.sub.3.sup.+, O, NH, NCH.sub.3, CH.sub.2 and CR.sub.1 R.sub.2 wherein at least one of R.sub.1 and R.sub.2 is not hydrogen; wherein each of Y.sub.2, Y.sub.3 and Y.sub.4 is selected from the group of N, NCH.sub.3.sup.+, CH, and CR wherein R is not hydrogen; and wherein at least two of Y.sub.1, Y.sub.2, Y.sub.3 and Y.sub.4 are neither CH.sub.2, CH, CR nor CR.sub.1 R.sub.2.

Abstract: Disclosed is a method for purifying Interleukin-10 (IL-10). The method is comprised of subjecting an IL-10 containing solution to cation exchange chromatography, anion exchange chromatography, hydroxyapatite chromatography, and gel filtration chromatography. The present invention is also comprised of a process for separating different IL-10 dimers present in a protein fraction from each other by subjecting the protein fraction to hydroxyapatite chromatography. The present invention is also comprised of a process for separating variants of a protein differing in an N-terminal amino acid sequence present in a protein fraction from each other by subjecting the protein fraction to hydroxyapatite chromatography.

Abstract: Methods and vectors for expressing interferon-alpha (IFN-.alpha.) proteins in E. coli are provided. Use of a vector comprising an IFN-.alpha. sequence fused to an E. coli heat-stable enterotoxin signal sequence (STII) under the control of the E. coli phosphatase (phoA) promoter affords high levels of correctly-folded and -processed, biologically active IFN-.alpha. polypeptides.

Abstract: A method for producing a stable polymerized hemoglobin blood-substitute from blood. The method of this invention includes contacting a polymerized hemoglobin solution with a hydroxyapatite HPLC column and recovering the substantially pure polymerized hemoglobin.

Abstract: Disclosed is a method for purifying Human interleukin-5 (IL-5) a single chromatographic step. After removing cells from a cell culture expressing human interleukin-5, IL-5 is purified by first adjusting the culture supernatant to the calculated pI value of mature IL-5 (pI=7.44) and then passing the conditioned supernatant through tandem linked anion- and cation-exchange columns. The resulting pass-through fraction contains the IL-5 and is devoid of all other contaminating proteins. An optional hydrophobic-interaction chromatography step is disclosed for positive selection of IL-5 and in order to concentrate the preparation. Pure IL-5 was recovered with a high overall yield (>90%), was N-glycosylated and was entirely homodimeric.

Abstract: In order to remove tumor necrosis factor .alpha. (TNF.alpha.) or/and bacterial lipopolysaccharides (LPS, endotoxin) extracorporeally from whole blood or/and blood plasma in an extracorporeal perfusion system, the blood or plasma is passed over a cation exchanger and an anion exchanger material. A device according to the invention for the extracorporeal treatment of patient's blood or plasma therefore contains a cation exchanger material and an anion exchanger material wherein these materials are contained in at least one compartment of an extracorporeal perfusion system.

Abstract: Disclosed is a process to remove tumour necrosis factor .alpha. (TNF.alpha.) or/and bacterial lipopolysaccharides (LPS) from an aqueous liquid, in particular blood, blood plasma or serum, in an extracorporeal perfusion system after removing corpuscular blood components if necessary, wherein(a) the pH value of the body fluid is adjusted to pH<6,(b) a precipitation reagent in the form of a polyanion is added,(c) precipitated substances are removed by filtration or/and centrifugation and(d) the resulting liquid is passed over an anion exchanger.

Abstract: Polyolefin particles are chemically modified by oxidation to provide a large surface area and high loading. The particles result in low back pressure in column systems, and are economical to manufacture. The particles are useful as supports in a wide range of applications including general organic as well as biopolymer synthesis, library methods, purification processes and enzyme mediated processes. In a preferred embodiment, polyethylene or polypropylene particles are oxidized in a solution containing trifluoroacetic acid or trifluoromethane sulfonic acid, chromium trioxide and sulfuric acid to provide the particles with a chemically reactive irregular surface and open channels that extend below the surface and up to essentially the length of the radius of the particles resulting in increased surface area and decreased density. The particles have pendant functional groups produced by the oxidation and/or by subsequent chemical reaction.

Abstract: The invention relates to a process for preparing a concentrate of Factor VIII-von Willebrand factor complex having high specific activity from total (non-cryoprecipitated) plasma.The process comprises pre-purifying by means of a double treatment with barium chloride and with aluminium hydroxide.The process then comprises purification by chromatography on an anion exchange resin, of the DEAE-Fractogel type.The process includes a step of viral inactivation by means of a treatment with solvent-detergent.The process also makes it possible to recover fibrinogen, albumin, immunoglobulins, antithrombin III, fibronectin and prothrombin complex, from the same plasma.The different concentrates obtained using the process according to the invention are intended, in particular, for therapeutic use.

Abstract: A cellulose binding domain (CBD) having a high affinity for crystalline cellulose and chitin is disclosed, along with methods for the molecular cloning and recombinant production thereof. Fusion products comprising the CBD and a second protein are likewise described. A wide range of applications are contemplated for both the CBD and the fusion products, including drug delivery, affinity separations, and diagnostic techniques.