Abstract: This invention relates to transgenically produced human Antithrombin III (tgATIII). The human ATIII produced by the transgenic process of the present invention has a monosaccharide composition which comprises N-acetylgalactosamine (GalNAc) along with fucose, N-acetylglucosamine, galactose, mannose, and N-acetylneuraminic acid/N-glycolyneuraminic acid. The monosaccharide composition differs with that of plasma derived ATIII (phATIII). It has been found that tgATIII has an increased clearance rate when compared to phATIII.

Abstract: The present invention relates to novel alternative forms of human acetylcholinesterase (AChE) and nucleotide sequences encoding the same. The genes encoding the novel forms of human AChE have been identified in various malignant tumor cells. In a further aspect, the invention relates to a transgenic animal assay system for evaluating efficacy of drugs against cholinergic proteins, prior to or in the course of therapeutic treatment. Transgenic animals, preferably developing tadpole of Xenopus or mice which express human AChE, are used. The transgenic animal assay system is also useful for evaluating the toxicity of substances which potentially block human AChE (e.g. organophosphorous compounds).

Abstract: A transgenic, non-human mammalian animal is capable of expressing a heterologous gene for human or other recombinant physiologically functional fibrinogen holoprotein or individual subunit chain polypeptides thereof or a modified or fusion fibrinogen in mammary glands of the animals and secreting the expressed product into a body fluid. Methodology employing such a mammal yields recombinant physiologically functional fibrinogens, subunit chain polypeptides thereof, and modified or fusion fibrinogens.

Abstract: Methods and processes for raising the concentration of a first class of immunoglobulin relative to at least a second class of immunoglobulin in a compartment of the body of a non-human animal or the progeny thereof, as well as the animals produced by such methods and processes. Such methods and processes provide for the collection of antibodies produced by mucosal surfaces of the animal. Preferably, the production is in the mammary gland. Antibodies can be collected from the milk of the animal. Antibodies may be used for medical and/or nutritional purposes.

Abstract: This invention relates to the production of recombinant proteins in mammals' milk. Particularly, this invention relates to an expression system which when transgenically incorporated into a mammal permits the female species of that mammal to produce the desired recombinant protein in or along with its milk. This invention also relates to the transgenic mammal that produces the desired recombinant product in its milk.

Abstract: The invention provides systems and methods for the production and purification of target molecules present in biological systems. The systems and methods according to the invention utilize transgenic expression of multivalent binding polypeptides, as affinity media, to purify such target molecules. The transgenic multivalent binding polypeptides bind both the target molecules, e.g., a bindable epitope of a target molecule, and a matrix.

Abstract: The invention provides systems and methods for the production and purification of target molecules present in biological systems. The systems and methods according to the invention utilize transgenic expression of multivalent binding polypeptides, as affinity media, to purify such target molecules. The transgenic multivalent binding polypeptides bind both the target molecules, e.g., a bindable epitope of a target molecule, and a matrix.

Abstract: Transgenically produced decorin and methods of making and using transgenically produced decorin.

Abstract: The invention provides transgenic nonhuman mammals producing phosphorylated lysosomal proteins in their milk, methods of generating the same, pharmaceutical compositions for use in enzyme replacement therapy, and methods of treating Pompe's disease using human acid alpha glucosidase.

Abstract: The present invention provides methods for the large-scale production of recombinant butyrylcholinesterase in cell culture, and in the milk and/or urine of transgenic mammals. The recombinant butyrylcholinesterases of this invention can be used to treat and/or prevent organophosphate pesticide poisoning, nerve gas poisoning, cocaine intoxication, and succinylcholine-induced apnea.

Abstract: The present invention encompasses a novel approach to reduce or to prevent modification of polypeptides in solution and to polypeptides obtained by such methods. Specifically, the invention relates to a method for reducing or preventing modification of polypeptides in milk, particularly milk obtained from a transgenic animal, and to polypeptides isolated using such methods.

Abstract: Production of human procollagen or collagen in cells which ordinarily do not produce these molecules is effected by constructing expression systems compatible with mammary glands of non-human mammals. For example, expression systems can be microinjected into fertilized oocytes and reimplanted in foster mothers and carried to term in order to obtain transgenic non-human mammals capable of producing milk containing recombinant human procollagen or collagen. Human procollagen or collagen produced in this manner can be made of a single collagen type uncontaminated by other human or non-human collagens.

Abstract: A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human FIX protein without the benefit of the presence of a complete milk gene sequence for gene rescue, and a signal sequence is active in directing newly expressed Factor IX into the milk of the animal at levels in an unactivated form that is suitable for subsequent processing for therapeutic applications in treating Hemophilia B. The transgenic mammals are preferably pigs, cows, sheep, goats and rebbits. The application include milk derivatives for oral delivery and oral tolerization in the treatment of Hemophilia B.

Abstract: A method of making a transgenic fusion protein. The method inlcudes providing a transgenic animal which includes a transgene which provides for the expression of the fusion protein; allowing the transgene to be expressed; and, recovering the fusion protein, from the milk of the transgenic animal.

Abstract: A non-human trangenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integratedintot he gemone of the animal, which comprises cis-acting regulatory untis operably linked to a DNA sequence encoding a modifed or variant human FVIII protein, and a signal sequence and secretion motif that is active in directing newly expressed Factor VIII into the milk of the animal at levels an in an unactivated, nondegraded and otherwise stable form that is suitable for subsequent processing for therapeutic applications in treating Hemophilia A. The transgenic mammals are preferably pigs, cows, sheep, goats and rabbits. The applications include milk derivatives used for oral delivery and oral tolerization in the treatment of Hemophilia A.

Abstract: The invention provides methods of treating Pompe's disease using human acid alpha glucosidase. A preferred treatment regime comprises administering greater than 10 mg/kg body weight per week to a patient.

Abstract: A DNA sequence containing a gene encoding a protein, the gene being under the transcriptional control in the DNA sequence of a mammalian milk protein promoter which does not naturally control the transcription of the gene, such DNA sequence including DNA enabling secretion of the protein.

Abstract: The present invention relates to animals that express exogenous growth factors in their milk, and in particular to pigs that express exogenous IGF-I in their milk. The present invention also relates to methods for increasing piglet weight gain and intestinal lactase activity. The present invention thus provides a method of facilitating piglet development and decreasing piglet mortality.

Abstract: The invention provides transgenic nonhuman mammals capable secreting exogenous procollagen or collagen into their milk. The mammals are healthy and capable of producing procollagen or collagen at high levels, usually in trimeric form. Suitable transgenes for incorporation into the mammals are also provided.

Abstract: The present invention provides methods for the large-scale production of recombinant butyrylcholinesterase in cell culture, and in the milk and/or urine of transgenic mammals. The recombinant butyrylcholinesterases of this invention can be used to treat and/or prevent organophosphate pesticide poisoning, nerve gas poisoning, cocaine intoxication, and succinylcholine-induced apnea.

Abstract: The present invention relates to animals that express exogenous growth factors in their milk, and in particular to pigs that express exogenous IGF-I in their milk. The present invention also relates to methods for increasing piglet weight gain and intestinal lactase activity. The present invention thus provides a method of facilitating piglet development and decreasing piglet mortality.

Abstract: A method for the production of monoclonal antibodies in mammal's milk, through the creation of transgenic animals that selectively express foreign antibody genes in mammary epithelial cells.

Abstract: The invention provides transgenic nonhuman mammals producing phosphorylated lysosomal proteins in their milk, and methods of generating the same. Phosphorylation occurs at the 6′ position of a mannose side chain residue. Also provided are methods of purifying lysosomal proteins from milk, and incorporating the proteins into pharmaceutical compositions for use in enzyme replacement therapy.

Abstract: Modified fusion proteins of transferrin and therapeutic proteins or peptides, preferably antibody variable regions, with increased serum half-life or serum stability are disclosed. Preferred fusion proteins include those modified so that the transferrin moiety exhibits no or reduced glycosylation, binding to iron and/or binding to the transferrin receptor.

Abstract: A milk or other dairy product, capable of minimising the onset of disease such as coronary heart disease or enhancing the immune response is derived from animals which are substantially free of the &bgr;-casein A1 allele. Bulk milk can be produced by testing for and culling cows who test positive for the &bgr;-casein A1 allele, or by producing immunoglobulins and other immune response proteins, in cow's milk from animals not possessing the &bgr;-casein A1 allele, or other commercial milk producing animals, to this allele, to counteract the immnunosuppressant substances present that are produced from it, in commercial milking cows such as Holsteins, together with its blending with non-treated milk or the recovery of such immunoproteins.

Abstract: A milk which is free of &bgr;-casein A1 protein in the prevention or treatment of coronary heart disease is disclosed. In addition, a process for the testing of DNA from cells obtained from lactating bovines for the presence of DNA encoding certain &bgr;-casein proteins, selecting the bovines on the basis of the testing, and then milking those bovines to produce milk free of &bgr;-casein A1 for use in the prevention or treatment of coronary heart disease is disclosed.

Abstract: Modified fusion proteins of transferrin and therapeutic proteins or peptides including soluble toxin receptors, with increased serum half-life or serum stability are disclosed. Preferred fusion proteins include those modified so that the transferrin moiety exhibits no or reduced glycosylation, binding to iron and/or binding to the transferrin receptor.

Abstract: A transgenic non-human animal expressing heparanase from a transgene, methods for its preparation, compositions-of-matter derived therefrom and uses thereof.

Abstract: The invention provides a method of making and secreting a non-secreted protein. The method includes expressing the protein from a nucleic acid construct which includes: (a) a mammary epithelial specific promoter; (b) a milk protein specific signal sequence which can direct the secretion of a protein; (c) optionally, a sequence which encodes a sufficient portion of the amino terminal coding region of a secreted protein to allow secretion in the milk of a transgenic mammal, of the non-secreted protein; and (d) a sequence which encodes a non-secreted protein, wherein elements (a), (b), optionally (c), and (d) are preferably operatively linked in the order recited. Both glutamic acid decarboxylase (GAD) and myelin basic protein (MBP), which are cytoplasmic proteins, have been produced by the methods of the present invention. The invention also provides methods for treating diabetes and multiple sclerosis using proteins produced by the methods of the present invention.

Abstract: A method of enhancing the transcription of a gene in a DNA construct incorporated into the genome of a eucaryotic host cell, wherein the DNA construct comprises a structural gene for a desired protein or polypeptide in a gene promoter upstream of the structural gene, comprises providing at least one enhancer element comprising the nucleotide sequence TTC TGA GAA upstream of the promoter, and exposing the DNA construct to lactogenic stimuli. An expression vector, a host cell and a transgenic mammal containing the vector are of interest.

Abstract: A process for the production of a peptide is disclosed, the process comprising expressing in the milk of a transgenic, non-human, placental mammal a fusion protein which comprises the peptide to be expressed linked to a fusion partner protein which is lysozyme. The fusion protein may be separate from the milk and cleaved to yield the target peptide. A transgenic, non-human, placental mammal whose genome incorporates a DNA molecule comprising a coding sequence encoding lysozyme coupled to a peptide is also described.

Abstract: A method of producing a substance comprising a peptide, involves incorporating a DNA sequence coding for the peptide into a gene of a mammal (such as a sheep) coding for a milk whey protein in such a way that the DNA sequence is expressed in the mammary gland of the adult female mammal. The substance may be an (optionally modified) protein such as a blood coagulation factor. The DNA sequence is preferably inserted into the first exon of a gene coding for a whey protein such as beta-lactoglobulin. The substance will generally be recovered from milk of the female mammal, but may (for example if it is an enzyme) be used in situ.

Abstract: A method and composition for administering leptin to a subject. The invention includes suspending isolated native leptin-containing milk fat globules in a suitable medium for administering to a subject. The suspended milk fat globules may be administered orally as well as by intravenous, intramuscular, intraperitoneal, other enteral routes of administration, and other parenteral routes of administration. The invention includes a method for treating growth or maturational-related disorders in newborns as well as subjects having conditions that can be treated by the administration of leptin.

Abstract: Proteinaceous molecules involved in molecular binding are disclosed. The binding molecules have a common structural core element that is provided with a desired affinity region. The core element and the desired affinity region can be adapted to bind to a wide range of molecules for a desired purpose. A method for identifying a binding molecule with an altered binding property and producing a nucleic acid for the identified binding molecule are also disclosed.

Abstract: The present invention relates to the stabilisation of milk from transgenic animals. In particular, the invention relates to the protection of proteins (e.g. fibrinogen) expressed in milk from transgenic animals by co-expression of a serine proteinase inhibitor (e.g. &agr;1-antitrypsin) in the milk of the transgenic animals.

Abstract: Described is a method for the production of a heteromeric protein comprising a first and a second peptide chain, wherein the first peptide chain is expressed in an animal, and the second peptide chain in another animal. The peptide chains are isolated from the animals and combined, resulting in formation of a functional protein. Optionally, both animals are mated, leading to offspring animals expressing both first and second peptide chains and producing the protein.

Abstract: The present invention provides a process for producing immunoglobulin A in the milk of hyperimmunised ruminants using a 3 route immunisation protocol and uses for the resultant product. These products are useful in producing formulations useful for passive immunisation against selected pathogens, especially in preparations for food products and dietary preparations.

Abstract: Proteinaceous products can be produced by transgenic animals having genetic constructs integrated into their genome. The construct comprises a 5′-flanking sequence from a mammalian milk protein gene (such as beta-lactoglobulin) and DNA coding for a heterologous protein other than the milk protein (for example a serin protease such as alpha1-antitrypsin or a blood factor such as Factor VIII or IX). The protein-coding DNA comprises at least one, but not all, of the introns naturally occurring in a gene coding for the heterologous protein. The 5′-flanking sequence is sufficient to drive expression of the heterologous protein.

Abstract: Transgenically produced prolactin and methods of making and using transgenically produced prolactin.

Abstract: The invention features transgenically produced PDGF, e.g., transgenically produced PDGF which is expressed in the milk of a transgenic mammal, and is present in the milk in active form, e.g., as a dimer. The invention also features methods of producing transgenic PDGF, transgenic animals capable of expressing PDGF, and nucleic acid sequences encoding PDGF, e.g., nucleic acid sequences encoding PDGF under the control of a mammary gland specific promoter.

Abstract: The invention provides a method of making and secreting a non-secreted protein. The method includes expressing the protein from a nucleic acid construct which includes: (a) a mammary epithelial specific promoter; (b) a milk protein specific signal sequence which can direct the secretion of a protein; (c) optionally, a sequence which encodes a sufficient portion of the amino terminal coding region of a secreted protein to allow secretion in the milk of a transgenic mammal, of the non-secreted protein; and (d) a sequence which encodes a non-secreted protein, wherein elements (a), (b), optionally (c), and (d) are preferably operatively linked in the order recited. Both glutamic acid decarboxylase (GAD) and myelin basic protein (MBP), which are cytoplasmic proteins, have been produced by the methods of the present invention. The invention also provides methods for treating diabetes and multiple sclerosis using proteins produced by the methods of the present invention.

Abstract: The present invention relates to human bile salt-stimulated lipase (BSSL) obtainable from transgenic sheep. The invention further relates to transgenic sheep whose germ cells and somatic cells contain a recombinant nucleotide molecule comprising a nucleotide sequence encoding for human BSSL. The invention also relates to methods for producing said transgenic animals, as well as to methods for producing human BSSL derived from transgenic animals. In addition, the invention provides the use of compositions comprising BSSL in the treatment of diseases relating to exocrine pancreatic insufficiency, and for improvement of the utilization of dietary lipids in preterm born infants.

Abstract: The invention features methods of purifying human serum albumin (hSA) from endogenous serum albumin of the host cell producing the hSA. The methods include providing a sample comprising hSA and serum albumin of the host cell, applying the sample to an affinity column that binds hSA at a higher affinity than the serum albumin of the host cell, eluting bound hSA from the affinity column, and crystallizing the eluted has. The invention also features compositions comprising hSA produced by the methods of the invention.

Abstract: Production of proteins not normally secreted through conventional pathways such as membrane proteins including, for example, CFTR associated with cystic fibrosis, is now made possible by collection of such protein from the milk of lactating transgenic animals.

Abstract: A non-human transgenic mammalian animal, as described above, contains an exogenous double stranded DNA sequence stably integrated into the genome of the animal, which comprises cis-acting regulatory units operably linked to a DNA sequence encoding human Factor VIII protein and a signal peptide, where the cis-acting regulatory units are active in mammary gland cells and the signal peptide is active in directing newly expressed Factor VIII into the milk of the animal. The promoter may be a milk protein promoter such as for whey acidic protein, casein, lactalbumin, or beta-lactoglobulin promoter. The transgenic mammals are preferably farm animals, for example, cows, goats, sheep, rabbits and pigs. Concurrent expression of a gene for human von Willebrand's Factor into milk may be used to stabilize newly-secreted Factor VIII.

Abstract: The invention features methods of making transgenic animals, and transgenic animals made by such methods. The method includes introducing into a cell, a nucleic acid construct comprising a nucleic acid sequence encoding a heterologous polypeptide under the control of a mammary epithelial cell promoter, an insulator positioned 5′ from the promoter, an insulator positioned 3′ from the nucleic acid sequence encoding the polypeptide, and a prokaryotic sequence, wherein the sequence between the insulator 5′ from the promoter and the insulator 3′ from the nucleic acid encoding the polypeptide is substantially free of prokaryotic sequence; and allowing a transgenic mammal to develop from the cell, to thereby provide a transgenic mammal.

Abstract: A milk composition that includes a heterologous non-milk allergen is administered to a subject to suppress allergen-specific IgE production in the subject

Abstract: A transgenic animal expressing heparanase from a transgene, methods for its preparation, compositions-of-matter derived therefrom and its uses.

Abstract: Process for producing mammals with defined genetic properties, particularly transgenic mammals. Animals which are derived completely from ES cells are obtained by injecting totipotent cells (embryonic stem cells or embryonic germ cells) genetically manipulated and cultured in vitro into tetraploid blastocysts and implanting the resulting embryo in a foster mother, in a single operation.

Abstract: Recombinant Factor IX characterized by a high percentage of active protein can be obtained in the milk of transgenic animals that incorporate chimeric DNA molecules according to the present invention. Transgenic animals of the present invention are produced by introducing into developing embryos DNA that encodes Factor IX, such that the foreign DNA is stably incorporated in the DNA of germ line cells of the mature animal. Particularly efficient expression was accomplished using a chimeric construct comprising a mammary gland specific promoter, Factor IX cDNA that lacked the complete or any portion of the 5′-untranslated and 3′-untranslated region, which is substituted with a 5-′ and 3′- end of the mouse whey acidic protein gene. In vitro cell cultures of cells explanted from the transgenic mammal of the invention and methods of producing Factor IX from such said culture and methods of treating hemophilia B are also described.