Abstract: An improvement is provided in a process for the membrane filtration of a protein solution. The improvement consists of applying a high shear force at the surface of the membrane. In embodiments of such process, the liquid is subjected to membrane filtration utilizing a rotating membrane disc mounted in close proximity e.g., about 1/8" to about 1/4" to a stationary solid disc, or a rotating solid disc mounted in close proximity e.g., about 1/8" to about 1/4" to a stationary membrane surface with the relative rotation being between bout 1,000 and about 3,450 RPM. This results in permeation characteristics of the selected membrane which essentially represents its theoretical molecular weight cut-off.

Abstract: A process is described for producing M-CSF from bacteria. It includes: fermentation of bacteria containing M-CSF DNA; harvest of the fractions that contain the M-CSF protein (refractile bodies); primary recovery of the protein; solubilization and denaturation of refractile bodies; M-CSF refolding; purification by column chromatography and other methods; and formulation of the properly refolded M-CSF. This method is advantageous over prior methods in terms of yield and purity.

Abstract: A process is provided for cleaving a polypeptide into at least two polypeptide components comprising treating a reduced, free-cysteine form of the polypeptide with a cleaving agent under conditions for cleaving the polypeptide at a desired junction between the polypeptide cleavage products. More preferably, the process for cleaving comprises culturing cells containing DNA encoding said polypeptide, wherein at least one Asp codon is present in said DNA at a desired junction between the components to be cleaved from each other, said culturing resulting in expression of the DNA to produce the polypeptide in the host cell culture; and treating a reduced, free-cysteine form of the polypeptide with dilute acid under conditions for cleaving the polypeptide at the Asp junction.

Abstract: A method for recovering hepatitis B surface antigen protein from transformed yeast cells including the steps of (i) obtaining an aqueous homogenate of the yeast cells; (ii) enriching the hepatitis B surface antigen protein in the homogenate with a protein-aggregating reagent to form a precipitate which contains hepatitis B surface antigen protein; (iii) dissolving the precipitate in a buffer to form a suspension; and (iv) post-homogenizing the suspension to obtain a 10% to 50% increase in yield of the hepatitis B surface antigen protein as calculated based on a yield achieved without performing the post-homogenizing step.

Abstract: Ala-Glu-IGF-I is a novel compound which exerts IGF-I activity and is a precursor for the preparation of IGF-I. Ala-Glu-IGF-I may by converted to IGF-I by renaturation after recombinant production in E. coli under specified conditions and then cleaving Ala-Glu from the IGF-I.

Abstract: An insoluble mammalian protein is extracted from transformed bacteria expressing the mammalian protein while avoiding irreversible insolubilization of bacterial host proteins by homogenizing the fermentation broth, centrifuging the homogenized broth and removing the supernatant liquid for the inclusion body containing pellet. In another embodiment, the pH of the homogenized broth is adjusted to 2.0 prior to centrifugation. The acidified broth is then centrifuged, and the pellet is resuspended in buffer, homogenized again, and the inclusion body is isolated by centrifugation.

Abstract: A carboxy terminal protein sequencing process is disclosed which utilizes phosphoroisothiocyanatidate for the derivatization step.

Abstract: TF55 is a homooligomeric complex of two stacked rings, closely resembling the quaternary structure of the chaperonins, groEL, hsp60, and RUBISCO-binding protein. Most rings of TF55 contain 9 radially arranged members. The TF55 complex binds unfolded polypeptides in vitro, preventing aggregation at elevated temperature, and exhibits ATPase activity, features consistent with its function as a molecular chaperone. At the level of primary structure, TF55 is not significantly related to the chaperonins but is highly homologous (36-40% identity) to a ubiquitous eukaryotic protein, t complex polypeptide 1 (PCT1).

Abstract: Tertiary amides have been developed which have low volatility and good thermal stability and which are effective in stabilizing organic materials that are normally susceptible to oxidative deterioration. The novel antioxidants are tertiary amides corresponding to the formula: ##STR1## wherein R is a mono-, di-, or trivalent aromatic or saturated aliphatic hydrocarbon group containing 1-20 carbons; n is an integer of 1-3 which is equal to the valence of R; R' is phenyl, benzyl, or C.sub.1 -C.sub.6 alkyl; m is an integer of 1-3; R" is a C.sub.1 -C.sub.4 alkylene group; Z and Z' are independently selected from hydrogen and alkyl; and Q is carbonyl or sulfonyl.

Abstract: Cryoprecipitated mammalian plasma proteins with associated glycoproteins, polysaccharides, and numerous other macromolecular entities are transferred directly in the course of controlled thawing and centrifuging from native plasma phase across the boundary layer into a pre-prepared substrate transfer medium at sustained solidus--liquidus equilibrium regulated to residual icing from about 5 weight percent to about 95 weight percent to produce cryoprecipitates with enhanced productivity and enhanced qualifications for in vivo tissue bonding applications.

Abstract: The protein doublet H110D, the individual components thereof, and the production and use thereof in a vaccine against a nematode infection. This protein doublet is a plasma membrane-associated protein material of the intestinal microvilli of Haemonchus contortus. H110D has a molecular weight of about 110 kd and reacts with antibodies raised in animals injected with a contortin-enriched fraction. Injection of preparations of the protein doublet H110D or its components induces the production of specific protective antibodies.

Abstract: The protein/starch bond is broken mechanically by wet attrition milling rather than by cooking or with chemicals alone. The grain particles are milled to a particle size sufficiently small to break the bond between starch and protein and sufficiently large to retain substantially all of the starch granules intact. The protein is then extracted with ethanol and alkali solvents, separated and dried to form protein and/or protein isolate. The intact starch granules are cleaned and dried.

Abstract: Disclosed is an improved method for hemoglobin purification utilizing a novel two-step dual-aqueous-phase extraction technique to separate hemoglobin from red cell membrane stroma and other protein contaminants. In the first step, a first dual-aqueous-phase liquid system is prepared which comprises an upper aqueous phase containing polyethylene glycol in water and a lower aqueous phase containing a phosphate buffer at a pH of about 10. In the second step, a second dual-aqueous-phase liquid system is prepared which comprises an upper aqueous phase, which contains the polyethylene glycol phase containing the hemoglobin extracted in the first step, and a lower aqueous phase containing a new phosphate buffer at a pH of less than 7.5. After the second extraction step, purified hemoglobin solution can then be obtained by removing the phosphate salt and the minute amounts of polyethylene glycol contained in the lower phase.

Abstract: Microwave heating of intact Trigonopsis variabilis cells to 60.degree.-68.degree. C. for 20-40 seconds selectively deactivates esterases and catalases while retaining the activity of D-amino acid oxidase. The heating is effected with a microwave frequency of 0.4-24 GHz.

Abstract: A process for facilitating the reconstitution of lyophilized Factor VIII complex compositions, and compositions of Factor VIII complex, which are readily reconstituted. The process of the present invention comprises providing a purified Factor VIII complex preparations; adding a stabilization agent comprising arginine; lyophilizing the stabilization agent-Factor VIII complex solutions; and reconstituting the lyophilized stabilization agent-Factor VIII complex by contacting it with solvent for less than one minute.

Abstract: Primary-toxic chemical compounds with a molecular weight of less than 12000 Daltons, which may be isolated from plant material and which are potential IgE-binding allergens causing immediate-type allergy in predisposed individuals; methods for their isolation and their use for clinical purposes.

Abstract: In the method for purifying factor VIII from cryoprecipitate, which is dissolved and then treated with alumina gel, the extract is diluted to a protein concentration not exceeding approximately 5 g/l and subjected to viral inactivation with solvent/detergent, the inactivated extract containing the solvent/detergent is then subjected to chromatography on a weak anion exchange column which is hydrophilic in nature and factor VIII is then eluted with a dissociating buffer.

Abstract: The present invention is a new nonantigenic keratinous protein material that may be used as a number of purposes, including correction of soft tissue deficiencies and the creation of biomedical implants and implant coatings. The present invention also includes processes for using the nonantigenic keratinous protein material for soft tissue augmentation, creating implants, and the coating of biocompatible implants. The nonantigenic keratinous protein material can be formed by obtaining nonantigenic keratinous protein and processing it to a powder form. If hair from the recipient or a compatible doner is used, it is bleached and rinsed, then dried and chopped into about 0.25 inch pieces. The keratinous protein is then homogenized in a solvent to a particular size generally in the range of about 0.1 to about 500 .mu.m. The particles are then ultrasonicated in a solvent.

Abstract: A vaccine for protecting avian and mammalian subjects against flea infestation comprises the supernatant fraction of flea midgut, or the antigenic components thereof. This also has the effect of reducing flea populations in the environment of the subject. Antibodies raised by these vaccines are also useful in purification and diagnosis.

Abstract: Proteins are prepared by the reaction of a hydrocarbon utilizing microbe on a low molecular weight hydrocarbon, such as methane, in an aqueous medium in the presence of oxygen and a fixable nitrogen compound in a recycle process in which unreacted hydrocarbon and oxygen are recycled to the protein manufacturing reactor. Sufficient hydrocarbon is introduced into the system to prevent the formation of a flammable mixture therein. Part or all of the hydrocarbon can be introduced into the system downstream of the protein reactor, where the propensity of forming a flammable gas mixture is greatest.

Abstract: A method for treatment of a matter which contains moisturized or liquified lactoferrin which has been isolated from mammalian milk, processed mammalian milk and by-products in mammalian milk-processing, without losing the physiological activities of lactoferrin, which comprises adjusting pH of said moisturized or liquefied lactoferrin contained in said matter within an acidic range between 2.0 and 6.0 both inclusive by adding acid or aqueous solution of acid when the pH of said moisturized or liquefied lactoferrin is out of said pH range, and heating said matter in the range from 60.degree. C. to 130.degree. C. for a span of time which may assure 60% or more of undenaturization rate of lactoferrin.

Abstract: The invention concerns a process for purifying protein A preparations to high purity with high product yield. Where the protein A is obtained from a Gram-negative recombinant microbe hosting a vehicle containing a gene encoding protein A, the protein A is purified to high purity, and, advantageously, to very low levels of endotoxin. The protein A preparations made via the invention process are useful in therapeutic application, e.g., therapeutic plasma exchange, as well as for other well-known uses of protein A.

Abstract: The present invention provides a method for purification of a surface exposed, immunogenic outer membrane protein of Haemophilus influenzas which is conserved amongst strains. The protein, designated P6, is relatively free of detergent, contaminating RNA and undesirable cellular components.In accordance with the present invention, there is provided a method for purifying an immunogenic outer membrane protein of H. influenzas consisting essentially of:a) suspending H.

Abstract: Processes for stabilizing active PAI-1 protein comprising combining active PAI-1 protein with an aqueous buffer having an ionic strength of at least about 5 millisiemens and a sugar selected from the group consisting of monosaccharides and disaccharides, and/or subjecting the active PAI-1 protein to lyophilization, are disclosed. Also described are compositions, including pharmaceutical compositions and kits, which include the stabilized active PAI-1 protein, and therapeutic processes employing the same in the treatment of fibrinolysis.

Abstract: The present invention relates to a process for purifying Factor IX from an impure protein fraction containing Factor IX. The purification process comprises the steps of adding a solvent and a detergent to an impure protein fraction and incubating the solvent/detergent protein solution to inactivate any viral contaminants. Factor IX is purified from the solvent/detergent protein solution by chromatography on a sulfated polysaccharide resin without first removing the solvent and detergent prior to the purification on the sulfated polysaccharide resin. The Factor IX, purified by the process has a specific activity of at least 85 units/mg.

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

Abstract: Methods of enhancing the dewatering of gluten are disclosed. The methods comprise adding a nonionic surfactant to the wet gluten prior to dewatering, as in vacuum dewatering equipment. Particularly effective surfactants are oxyalkylated sorbiton R surfactants when R is monooleate, trioleate, monostearate, tristearate, monopalmitate and monolaurate.

Abstract: A process is disclosed which makes possible the isolation of the luminal endothelial cell membrane from associated tissue. It is particularly applicable to vasculature, but broadly is applicable to all tissue cavities which are accessible from adjacent perfusable lumens. The method involves the identification of characteristic molecules (primarily proteins and lipids) associated with the luminal surface of the any endothelial membrane in situ by utilizing a novel membrane-isolation scheme to separate the endothelium from associated tissue. In this method, the endothelial luminal plasmalemma of a given organ is coated with colloidal silica by perfusion, a pellicle is formed, the coated area of tissue is excised and the coated plasmalemma fragments are isolated from the cognate homogenate by centrifugation. The isolated plasmalemma attached to the pellicle can then be subjected to biochemical analysis to identify and catalogue molecules characteristic of the endothelial membrane.

Abstract: A group of growth factors, designated heparin-binding brain mitogens (HBBMs), is disclosed. The HBBMs are isolated from brain tissue by a sequence of purification steps. The growth factors may be useful in the promotion of angiogenesis, such as in the promotion of wound healing, bone healing and in the treatment of burns, as well as in promoting the formation, maintenance and repair of tissue, in particular, neural tissue.

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

Abstract: A refractile material containing a heterologous protein is recovered from a host microorganism cell culture transformed to produce the protein. One recovery process involves first reducing the ionic strength of the culture medium prior to disruption to a level effective in preventing reaggregation of cellular debris with refractile material after disruption, disrupting the desalted culture, optionally adding material to the disruptate to create a density or viscosity gradient and separating the refractile material from the cellular debris by high-speed centrifugation. Preferably the salt removal step is carried out by diafiltration and the heterologous protein comprises recombinant M-CSF, IL-2 or IFN-.beta..

Abstract: Casings for food such as sausage are prepared from connective tissue that has been removed from animal tissue in accordance with the process comprising the steps of (1) mechanically separating connective tissue and impurities from animal tissue; (2) treating the connective tissue to remove the fat; (3) reducing the residual bone content of the connective tissue by exposure to acidic materials; (4) separation of the non-collagenous protein and elastin by treating the connective tissue with a first enzyme; and (5) separation of the muscle tissue from the connective tissue by treating with a second enzyme. The step of removing the fat from the connective tissue can be accomplished by one or more of the following steps: (A) hydrocarbon extraction of the fat from the connective tissue; (B) extraction of the connective tissue with a critical fluid; (C) dissolution of the fat with a third enzyme; (D) treatment of the fat with a suitable treating agent; (E) heat treatment; and/or (F) pressure treatment e.g.

Abstract: Crude immunoglobulin G isolated from human blood plasma is treated according to a conventional technique (such as the tricalcium phosphate adsorption method) to remove aggregates therefrom to such an extent that they are not detectable by gel filtration analysis. In order to produce an aqueous solution of immunoglobulin G having a reduced anticomplementary activity, the resulting solution is then filtered through a porous polyolefin membrane having a pore size larger than the molecular size of immunoglobulin G, in the presence of a stabilizer having surface activity. The aqueous solution of immunoglobulin G so produced is suitable for use in intravenous injection because its anticomplementary activity is low.

Abstract: The present invention relates to a dried composition containing IGF-I which is highly soluble and has a long shelf life stability. The present invention further relates to a method of preparing a dried composition containing IGF-I by drying a solution containing IGF-I together with a strong acid which is hydrochloric acid, hydrobromic acid, nitric acid, methanesulfonic acid, sulfuric acid, phosphoric acid, or oxalic acid.

Abstract: According to the process the sludge is homogenized and heated to 30.degree.-60.degree. C. temperature. The heated sludge is circulated under 2-4 bar overpressure, and its temperature is raised to about 130.degree.-150.degree. C. in 1-2 sec by conducting steam of 130.degree.-151.degree. C. and 2-4 bar pressure directly into the sludge, thus granulation is brought about in the sludge. The sludge of increased temperature--while its temperature and pressure maintained--is further circulated for about 60-300 sec. Then, the sludge is adiabatically expanded in 1-2 sec by reducing the pressure to about 0.01-0.02 bar. The expanded sludge is separated to water, fat containing water and solid impurity and wet solid phase containing protein.It is characteristic to the apparatus that it is provided with device for heating the sludge, and pipe (18) for passing on the heated sludge connected with the heating unit (5) where an expansion valve is built in.

Abstract: Particles having an average diameter of less than about 12 microns comprising fatty acid cores encapsulated with human serum albumin and methods for their preparation are disclosed. These materials are useful as contrast agents in ultrasonic imaging, having scattering intensities that are equivalent to or greater than those obtain from dispersed microbubbles but being much more stable, both in storage and when used in vivo, than are contrast agents based on dispersed microbubbles.

Abstract: A formulation for proteins is provided that comprises a lyophilized mixture of a protein, a water-soluble etherified cellulose in an amount that upon reconstitution of the formulation will be sufficient to form a gel, and an excipient to facilitate rehydration of the gel and to maintain protein integrity during storage of the lyophilized gel product. When reconstituted to a gel, this formulation can be applied, for example, to tissue in need of treatment.

Abstract: A method of making a biologically inactive polypeptide active is disclosed. Activity is imparted to the polypeptide through treatment with an exogenous peptide sequence. The nature of the exogenous peptide sequence is disclosed.

Abstract: A carboxy terminal protein sequencing process is disclosed which utilizes phosphoroisothiocyanatidate for the derivatization step.

Abstract: A stable immunogen composition for oral administration which includes a dried spherical form comprised of an immunogen capable of immunizing human or animals and a gelatin having an average molecular weight of 80,000-120,000 and jelly strength of more than 150 (Bloom, g, 6.2/3%), and is enteric.

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 composition containing a high concentration of a biologically active drug and a method for the use thereof to reduce vial breakage in a lyophilization process are disclosed.

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: The invention relaates to the production of collagen fibers by comminuting collagen containing tissues, drying the comminuted product and milling the dried material while maintaining the temperature sufficiently low to prevent substantial conversion of collagen to gelatin. The collagen fiber product is particularly useful for restructuring poorly textured meats, mechanically recovered meat products, offal, fish, fish products and other protein products to improve textural properties, water retention, fat retention, eating quality, juicines, succulence, shape, size retention and protein content.

Abstract: The chlorohydrin content of a liquid containing hydrolyzed protein obtained by hydrolysis of protein with hydrochloric acid is reduced in an apparatus system by introducing alkali into the liquid flowing under pressure in a piping system to increase the pH of the liquid, after which the pH-increased flowing liquid is heated, and then the heated flowing liquid is held for a time sufficient in a holding piping section to reduce the chlorohydrin content of the hydrolyzed protein contained in the liquid. The flowing liquid under pressure then is cooled before or after addition of hydrochloric acid which is employed to adjust the pH of the liquid.

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 process for extracting type I collagen from an avian source such as poultry feet that incorporates a fibrillar mass of connective tissue as well as bony tissue to yield a collagen product having useful medical and biotechnology applications. In this process, after being cleaned and decontaminated, the poultry feet are comminuted and then enzyme-treated to enhance the yield. The enzyme-treated comminuted material which is rich in collagen is dispersed in an organic acid to cause the fibrillar mass to undergo controlled swelling, after which the mass is separated from the bony tissue and purified to remove non-collangenous material. The purified mass is dried to provide the desired Type I collagen product which may be ground into a powder or formed into a collagen matrix or sponge, depending on the end use therefor.

Abstract: A high concentration of renaturing surfactants is added to protein systems in the presence of alkyl sulfate detergents to displace the detergent with respect to the interaction with the protein, thereby renaturing the protein and restoring its reactivity. The presence of higher quantities of detergent in the system results in smaller protein aggregates, and thus a higher reactivity and specificity as measured for the system in its entirety.

Abstract: A method of extracting granulocyte/macrophage colony stimulating factor (GM-CSF) from GM-SCF-expressing bacterial cells comprising treating a suspension of GM-CSF-containing bacterial cells with an acid and an enhancing agent, or with an acid that is itself an enhancing agent, removing substantially all of the suspension liquid from the cells, preparing a second suspension of the acidified cells, neutralizing said second suspension, and separating the GM-CSF-containing liquid from the suspended cells.

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.