Abstract: The present invention provides a process for the recovery of heterologous proteins from CTAP-III fusion proteins comprising expressing a fusion protein having a first amino acid sequence, a second amino acid sequence, and a selectable site which may be cleaved to provide first and second polypeptide fragments, respectively, wherein the first amino acid fragment is homologous to CTAP-III, and the first and second fragments have different pI values; cleaving the fusion protein to provide the first and second fragments; and separating the first and second fragments by ion exchange chromatography.

Abstract: A method of removing an organic solvent from a mixture that includes a compound having a polypeptide segment and the organic solvent, the method including the steps of contacting the mixture with an ion exchange resin under conditions that allow the compound to bond to the resin; and washing the resin with a first aqueous solution that elutes the organic solvent from the resin while allowing the compound to remain bound to the resin.

Abstract: Monoclonal antibodies specific for epitopes found on the B chain of PDGF (including v-sis, c-sis and platelet-derived forms) may be bound to columns and used for purification of rPDGF B. A solution containing a polypeptide possessing at least part of the structural conformation of rPDGF B is passed over such a column and the rPDGF B is bound to the antibody. The rPDGF B may then be eluted from the column to yield rPDGF B of greater than 95% purity as determined by SDS-PAGE.

Abstract: A process for the purification of proteins from solutions containing contaminants of similar net charge and molecular weight is provided, comprising contacting a solution containing the desired protein with an immobilized metal affinity chromatography resin in a buffer containing a low concentration of a weak ligand for the chelant of the resin. The adsorbed protein is then eluted using a buffer having a high concentration of the same weak ligand, e.g., Tris. Particularly preferred features employ agarose-iminodiacetic acid resins having copper cations and are especially useful in obtaining preparations of homogeneous, stable rsT4 proteins.

Abstract: Novel neutrotrophic factor compositions are provided, obtained from lung tissue. The factors are found to be active on parasympathetic ganglion neutrons enhancing acetylcholine activity. The compositions find use in the treatment of a number of eye disorders.

Abstract: Disclosed herein is a process for separation and analysis of free and bound hydrophobic components in whole blood. In this process, whole blood is passed through an internal surface reverse phase material having particles large enough to allow blood cells to pass through. These particles have hydrophobic pores that are small enough to prevent penetration by proteinaceous substances and large enough to allow penetration by free hydrophobic components. Thus, free hydrophobic components are retained by the internal pore surfaces. A non-denaturing solvent is used to wash the whole blood through the material, and a hydrophobic or organic solvent may be separately used to wash the hydrophobic components from the material. This process may be combined with qualitative, quantitative, and selective detection techniques, such as radiolabelling.

Abstract: A process for recovering highly pure, recombinant IL-2 from transformed microorganisms in which the cells are disrupted; impure recombinant IL-2 is isolated in the form of refractile bodies from the disruptate; the impure IL-2 is dissolved and denatured with at least 6M guanidine hydrochloride containing a reducing agent; the reduced IL-2 is precipitated and resolubilized; the reduced solubilized IL-2 therein is oxidized by a controlled oxidation; the oxidized IL-2 is refolded by reducing the concentration of guanidine hydrochloride in the solution; and the oxidized, refolded IL-2 is further purified by ion exchange chromatography or hydrophobic interaction chromatography and ion exchange chromatography.

Abstract: Methods are provided for the purification of interleukin-2 (IL-2) from a wide variety of sources, including synthetic mixtures, culture medium conditioned by natural IL-2 producing cells, and mammalian and bacterial recombinant IL-2 expression systems. The methods of the invention employ IL-2 receptor-affinity adsorbents in which soluble IL-2 receptors have been immobilized on solid supports. Through the use of these affinity adsorbents, highly purified IL-2 can be produced in a single step from bacterial extracts or conditioned medium. The IL-2 thus purified is largely free of aggregated forms, which are often present when other purification methods are used.

Abstract: An essentially pure and stablized antibody preparation comprising IgM antibodies having a purity greater than about 98% by weight and a nucleic acid content of less than about 200 pg per mg IgM. In one embodiment IgM antibodies from a monoclonal source are subjected to ion exchange and size exclusion chromatography at an alkaline pH to yield a purified IgM having a nucleic acid content of less than 10 pg/mg IgM, preferably less than about 4 pg/mg IgM. A highly purified and stabilized preparation of anti Pseudomonas aeruginosa antibodies is disclosed. The removal of nucleic acids is assured by subjecting the antibody source to at least one and preferably two low pH precipitation steps. In a very preferred embodiment, ion exchange and/or size exclusion chromatography is used to remove any residual nucleic acids.

Abstract: A megakaryocyte stimulatory factor (MSF), purified to homogeneity, is an acidic protein (pI=5.1) with an Mr=15,000 which stimulates PF4-like protein synthesis in rat promegakaryoblast cells by as much as 7-fold, and exhibits half-maximal activity at a concentration of 0.8 pM. MSF exhibits no biologic activity corresponding to other known hemopoietic growth factors, and appears to be specific for the megakaryocyte lineage.In the given examples, MSF was purified to homogeneity (as judged by SDS-PAGE and isoelectric focusing in the presence of 9.2 M urea) from serum-free conditioned medium obtained from cultured human embryonic kidney (HEK) cells, and to near homogeneity from thrombocytopenic plasma.

Abstract: Methods are provided for the purification of interleukin-2 (IL-2) and chimeric proteins containing an IL-2 moiety from a wide variety of sources, including synthetic mixtures, cell culture conditioned medium and mammalian and bacterial recombinant expression systems. The methods of the invention employ IL-2 receptor-affinity adsorbents in which soluble IL-2 receptors have been immobilized on solid supports. Through the use of these affinity adsorbents, highly purified IL-2 and chimeric proteins containing an IL-2 moiety can be produced in a single step from bacterial extracts or conditioned medium. The proteins thus purified are largely free of aggregated forms, which are often present when other purification methods are used.

Abstract: A recombinant DNA fragment (Z) coding for an immunoglobulin G binding domain related to staphylococcal protein A. The methionine codon of the recombinant DNA fragment has been replaced with a codon of an amino acid residue other than methionine to enable expression of a methionine-free protein. The present invention is also directed to recombinant DNA sequence containing such recombinant DNAS fragment (Z) as well as a recombinant DNA molecule, plasmid vector and bacterial cell containing such recombinant DNAS fragment. Also disclosed is a process for cleaving a fused protein expressed in a biological system using said recombinant DNA fragment.

Abstract: The present invention provides cleavable conjugates whose linkers contain a labile bond that is cleavable under a variety of mild conditions, including weakly acidic. Since the agent may be bonded directly to the linker, cleavage can result in release of native agent. The invention also provides methods for producing cleavable conjugates. Preferred agents include drugs, toxins, biological response modifiers, radiodiagnostic compounds, radiotherapeutic compounds, and derivatives thereof. The targeting molecule employed in the invention may be an intact molecule, a fragment thereof, or a functional equivalent thereof. In a particularly preferred embodiment, the targeting molecule is a monoclonal antibody directed towards a tumor-associated antigen in man. The invention further provides methods for delivering to the cytoplasm of a target cell an agent free of its targeting molecule carrier.

Abstract: A microporous structure with layered interstitial surface treatments, and method and apparatus for preparation thereof. The structure is prepared by sequentially subjecting a uniformly surface-treated structure (10a) to atomic oxygen treatment to remove an outer layer (16) of surface treatment to a generally uniform depth, and then surface treating the so exposed layer with another surface treating agent. The atomic oxygen/surface treatment steps may optionally be repeated, each successive time to a lesser depth, to produce a microporous structure having multilayered surface treatments. The apparatus (200) employs at least one side arm (228) from a main atomic oxygen-containing chamber (202). The side arm (228) has characteristic relaxation times such that a uniform atomic oxygen dose rate is delivered to a specimen (239) positioned transversely in the side arm (228) spaced from the main gas chamber (202).

Abstract: A novel and unique plasminogen activator inhibitor fragment is obtained from human umbilical vein endothelial cells which has the following characteristics:A. it is derived from a native t-PA inhibitor that binds to and inhibits the activity of t-PA,B. it is dissociated from a complex formed between said native t-PA inhibitor and t-PA, said complex existing in two distinct interconvertible conformations with molecular weight of about 88 KDa and 105 KDa, respectively, and being partially reversible in the presence of fibrin,C. it has a molecular weight of about 40 KDa when dissociated from the complex, andD. it has a novel partial N-terminal amino acid sequence when dissociated from the complex.

Abstract: Magnetically stabilized fluidized bed technology is utilized in conjunction with ion-exchange adsorption/desorption processes in a method and system for isolating proteins from cell lysate. The invention also includes a magnetizable, porous, ion-exchange particle, and a method for producing the same, for use with the stationary magnetically stabilized fluidized bed protein isolation process.

Abstract: An antibacterial polypeptide is disclosed, which is immunologically produced by Sarcophaga peregrina, when the insect is injured in its body wall. The polypeptide has a molecular weight of about 7000.

Abstract: Improved methods for direct purification of product immunoglobulins or their derivatives from large volumes of mammalian cell culture medium include directly subjecting the cell culture medium to cation exchange treatment, so as to adsorb the product but not the contaminants. The eluted product is then recycled, or is applied to anion exchange, for further purification, and optionally subjected to additional steps. The product may be obtained in a form suitable for clinical applications, if desired.

Abstract: Substantially pure proteins active in humoral hypercalcemia of malignancy (PTHrP) and sub-units and fragments thereof. Antibody reagents capable of binding to epitopes of PTHrP. Methods and kits for the detection of PTHrP.

Abstract: Antibody preparation purified using immobilized protein A and yet substantially free of protein A that may have solubilized during the purificaiton process. The antibodies include less than 15 ng protein A per mg of antibody, preferably less than 1 ng/mg. Low protein A content is obtained by first contacting the antibodies and solubilized protein A with an ion exchange resin under conditions to adsorb both. The antibodies and protein A are then sequentially eluted under conditions of increasing ionic strength.

Abstract: A method of and apparatus for separating proteins adsorbed by ion-exchange gels, especially anion-exchanging gels includes at least one chromatographic column in which the protein-carrying gel is charged and is subjected to a gradient-elution with a buffer solution as eluant whose property is changed with time by gradually changing the ionic strength and maintaining a substantially constant pH value or by gradually changing the pH value and maintaining substantially a constant ionic strength. The obtained eluate is then fractionated into its various components.

Abstract: The present invention provides a method for the separation of anti-metal chelate antibodies from non-specific proteins, including antibodies, by applying a preparation containing the anti-metal chelate antibodies to an oxo acid derivatized solid support and eluting first with an elution buffer containing sufficient salt concentration to elute non-specific proteins but not sufficient to elute the anti-metal chelate antibodies and then increasing the salt concentration of the elution solution so as to elute the anti-metal chelate antibodies. In one embodiment, the oxo acid derivatized solid support is a sulfopropyl resin. Appropriate salts include sodium phosphate, sodium chloride and sodium acetate. The method can be used to separate monoclonal or polyclonal anti-metal chelate antibodies from non-specific proteins as well as to separate bifunctional anti-metal chelate antibodies from monoclonal anti-metal chelate antibodies and other non-specific proteins.

Abstract: A method is described for safely and effectively washing solid gels or films containing biological macromolecules such as proteins or nucleic acids. The method involves placing the gels in an open plastic container and decanting a wash solution horizontally so as to leave the gel at the bottom of the container without using external means to hold the gel down. The undamaged gel can then be safely removed and detection experiments can thus be carried out. A preferred apparatus for carrying out this method is also described and consists of an elongated open-topped side spouted tray.

Abstract: A method for purifying from a mixture of hybrid compound which contains a portion of IL-2, which portion includes at least a region of the IL-2R binding domain of IL-2, which region is effective to cause the hybrid compound to bind to cells containing an IL-2R. The method includes passing the mixture containing the hybrid compound over a column having attached thereto molecules consisting of or containing a complex sugar moiety with affinity for the hybrid compound, and eluting the hybrid compound with a suitable eluant. The invention also features related complexes of the hybrid compound and assays for the hybrid compound.

Abstract: The disclosure describes novel immunosuppressive agents isolated from syncytiotrophoblast microvilli membranes by preparing a minutely subdivided and solubilized preparation of said membranes and isolating the unreduced N-linked oligosaccharides from an extract of the preparation.

Abstract: A process for obtaining a material possessing glucose tolerance factor activity and capable of binding with insulin including the steps of (i) contacting a eukaryotic cell mass with a mixture of water and alcohol to form a water phase and an alcohol phase; (ii) separating the alcohol phase from the water phase and isolating a water phase extract; and (iii) subjecting the water phase extract to gel exclusion chromatography, eluting all material having a molecular weight of approximately 720 to 1120 to obtain a material possessing glucose tolerance factor activity and capable of binding with insulin.

Abstract: This invention provides a crystalline form of recombinant human granulocyte-macrophage colony-stimulating factor (r-h-GM-CSF) and methods for making such crystals.

Abstract: Recombinant hepatitis B virus surface proteins produced in recombinant host cells are rapidly and efficiently purified from either cell extracts in a high pH buffer, or from heated whole cells at neutral pH. The host cell extracts or whole cells are heat treated, cooled and in the case of high pH extract, the pH is reduced. The surface proteins are then absorbed onto wide pore silica followed by elution and concentration. This method eliminates the requisite introduction of protease inhibitors, stabilizes the surface protein and improves product yield.

Abstract: A protein which satisfies all the biological criteria which are characteristic of inhibin has been isolated from a gonadal source. The purification and characterization of inhibin and the use of the purified material to raise antibodies, the use of inhibin and said antisera in a quantative radioimmunoassay, and applications in vitro and in vivo of inhibin and antibody to inhibin, are described.There is provided a purified protein, inhibin, characterised in thata. the apparent molecular weight as determined by SDS-PAGE is 56,000.+-.1,000b. the isoelectric point is in the range 6.9-7.3c. the protein can bind specifically to Concanavalin A-Sepharosed the protein consists of two sub-units, characterized in thati. their apparent molecular weights as determined by SDS-PAGE are 44,000.+-.3,000 and 14,000.+-.2,000 respectively.ii. the isoelectric point of the 44,000 molecular weight sub-unit is in the range 6.0-7.0iii. the N-terminal amino acid sequences of the two sub-units are as described hereine.

Abstract: The present invention comprises a method for the purification of the 69 kDa outer membrane protein of Bordetella B. pertussis and the protein purified therewith. A preferred embodiment comprises the purification of the 69 kDa protein from Bordetella B. pertussis strain Bp 353. The present process is advantageous in that it does not require or involve the use of biologics (such as monoclonal antibodies) and therefore simplifies the purification procedure and makes the resulting purified protein particularly advantageous for inclusion in acellular vaccines.

Abstract: Basic proteins are isolated from protein mixtures which contain such basic proteins and which are obtained by enzymatic clevage of proinsulin and/or its derivatives of natural, semisynthetic or genetic engineering origin by ion exchanger chromatography on strongly acid cation exchangers.

Abstract: An agent for the therapy or prophylaxis of disturbances of hemostasis, which contains tissue protein PP4, a process for the purification of PP4 by means of a hydrophobic adsorbent which is insoluble in water, and a process for the pasteurization of a solution containing PP4 in the presence of calcium ions and of at least one mono- or oligosaccharide or sugar alcohol and, where appropriate, of an amino acid, and the use of PP4, are described.

Abstract: The invention relates to a process for the isolation and purification of hirudin from complex and salt-containing solutions by hydrophobic chromatography, using as stationary phase porous adsorber resins and as mobile phase organic solvents which are miscible with water.

Abstract: The gene encoding protease nexin I (PN-I) is cloned and expressed to provide practical quantities of PN-I for diagnostic and therapeutic use. PN-I is a serine protease inhibitor useful in controlling conditions mediated by proteolytic activity.

Abstract: Disclosed is a method for separating and recovering very small amounts of biologically active proteins (such as enzymes, antigens and antibodies) present in a various fluids, with a high recovery and on a large scale, in which a fluid containing a desired protein is introduced into a rotary column through orifices therein and brought into contact with a gel carrier capable of selectively adsorbing the desired protein, while the carrier is being gentle fluidized in a liquid by the rotary motion successively by wash water and eluting fluid, so that the column, after which the fluid is displaced, the carrier having the desired protein adsorbed thereon is washed and the desired protein is recovered therefrom by elution while the carrier fluidized.

Abstract: A method for separating organic compounds, specifically proteins, is provided. The method for separating organic compounds is based on a combined pi-electron and electrostatic interaction of the exchanger with specific components of the organic compound. The method provides both an anionic and cationic exchanger. Each exchanger can provide more than one dimension of separation.

Abstract: A blood substitute and plasma expander comprising a cross-linked, substantially endotoxin-free homoglobin solution and process for preparing same. The process comprises fractionating whole blood, separating out a stromal-free, sterile hemoglobin solution, chromatographically separating endotoxins from said hemoglobin solution and crosslinking the resulting endotoxin-free hemoglobin solution.

Abstract: A new method for the purification of a 168 kD protein from Mycoplasma pneumoniae using zwitterionic and non-ionic detergents.

Abstract: A solid support useful for bioaffinity and ion-exchange separations and enzyme immobilization is provided. The support is based on a non-perfluorocarbon solid carrier core coated with a nonionic fluorosurfactant-coated fluorocarbon interlayer to which a ligand or a binder for the ligand is securely, but reversibly attached through a reactive perfluorocarbon anchor compound. Also provided is a solid support useful for size exclusion separations. Such support is based on a non-perfluorocarbon carrier core coated with a nonionic fluorosurfactant-coated fluorocarbon interlayer.

Abstract: The invention relates to a process for the preparation of purple membrane containing bacteriorhodopsin, which process comprises obtaining, in a manner know per se, the cell membrane from halobacteria cells, and subjecting the material to gel filtration chromatography in order to isolate the purple membrane from the cell membrane.

Abstract: This invention deals with a simple and efficient method for producing vitronectin from biological materials which include vitronectin. More specifically, this invention deals with a method for producing vitronectin by binding vitronectin in biological materials to immobilized glycosaminoglycans in the presence of protein denaturing agent, especially urea.

Abstract: A substantially pure, hemagglutinin-free composition of trichosanthin, and a method of producing the composition is disclosed. A plant extract from Trichosanthes kirilowii is contacted with an anionic exchange resin to remove contaminating hemagglutinins, and trichosanthin is further purified from the extract by cation exchange chromatography to a purity of greater than 95%.

Abstract: New lymphokine and its isolation and purification process. Said lymphokine is comprised of a factor obtained from T cells stimulated by concanavaline A or by an antigen capable of inhibiting the IgE-dependent platelet citotoxicity with respect to young larvae of S. Mansoni, of strongly reducing the chemiluminescence of blood platelets in a reaction IgE-anti-IgE, which is a correlate of the anti-parasite cytotoxicity, and of inhibiting the platelet activation in non-IgE dependent intolerences. Application as suppressor agent for suppressing platelet activation and as immunomodulator medicament of allergies.

Abstract: A method is described for the purification of crude human interferon from solutions containing it, which comprises:a) the complete adsorption of the crude interferon in a column of siliceous material which has previously been disinfected with an aqueous solution of formaldehyde;b) the washing of the column with non-pyrogenic, sterile, deionized water;c) the removal of the extraneous residual proteins by the elution of the column successively with a 1.4 M aqueous solution of NaCl in non-pyrogenic, sterile, deionized water, and with an aqueous solution of acetic acid having a molar concentration of 0.001 M to 0.003 M;d) the elution of the interferon from the column with an aqueous solution of acetic acid having a molar concentration of from 0.01 to 0.03 M and finally,e) the recovery and lyophilization of the elution containing the purified interferon.

Abstract: A protein containing at least one pendant sulfhydryl group is directly radiolabeled with a radiometal which binds tightly to sulfhydryl groups, using one or more pendant sulfhydral groups on the protein as endogenous ligands and optionally using an exogenous ligand which binds tightly to the radiometal ion to further stabilize the chelate.

Abstract: A process for purifying dithiochrome from a natural source is disclosed. The process comprises: (i) contacting a eukaryotic cell mass with a mixture of water and an alcohol; (ii) separating the alcohol phase from the water phase and obtaining a water phase extract; (iii) subjecting the water phase extract to a sulfhydryl exchange chromatography matrix; (iv) washing said chromatography matrix; and (v) treating said chromatrography matrix with an active thiol to obtain dithiochromate; a material capable of binding with insulin. Dithiochromate may be used to treat patients suffering from elevated blood glucose or suboptimal glucose kinetics. An adduct of this compound with insulin may be used to treat patients for type I diabetes.

Abstract: A preparation of an isolated immunoreactive CA 125 Antigen, and a method of isolating it is disclosed. CA 125 Antigen is a glycoprotein having a molecular weight of about 200kD, and a carbohydrate-content of about 24%. The CA 125 Antigen is isolated from a cell culture medium by acid precipitation, and is subsequently purified by size exclusion chromatography and immunoaffinity chromatography.

Abstract: A process is disclosed for the purification of Nerve Growth Factor (.beta. subunit) by subunit exchange chromatography.

Abstract: A process for purifying human G-CSF from an aqueous solution including the step of adding NaCl to the solution to selectively precipitate the human G-CSF.

Abstract: A homogeneous composite polymer containing, in an interpenetrated form, 20% to 80% by weight of silica and 80% to 20% by weight of a three-dimensional crosslinked acrylic, vinyl and/or allyl copolymer comprising, in a copolymerized form, 98% to 70% by weight of at least one monofunctional acrylic, vinyl or allyl monomer and 2 % to 30% by weight of a difunctional acrylic or allyl crosslinking monomer is useful in liquid chromatography.