Abstract: Apparatus and methods are described for pharmaceutical grade manufacture extrachromosomal nucleic acids from cell lysates using flotation to separate and eliminate undesired insoluble cellular debris including chromosomal DNA from the lysates. A gas is introduced to controllably generate bubbles that reduce the density of the cell debris and create a buoyant flocculent phase that can be readily separated from, and thus provide, a substantially clarified fluid lysate phase that is enriched in extrachromosomal DNA but substantially depleted of cellular proteins and chromosomal DNA.

Abstract: Apparatus and methods are described for pharmaceutical grade manufacture extrachromosomal nucleic acids from cell lysates using flotation to separate and eliminate undesired insoluble cellular debris including chromosomal DNA from the lysates. A gas is introduced to controllably generate bubbles that reduce the density of the cell debris and create a buoyant flocculent phase that can be readily separated from, and thus provide, a substantially clarified fluid lysate phase that is enriched in extrachromosomal DNA but substantially depleted of cellular proteins and chromosomal DNA.

Abstract: A scalable alkaline lysis process, including procedures and devices for the isolation of large quantities (grams and kilograms) of plasmid DNA from recombinant E. coli cells. Effective, controllable, and economical operation, and consistent low level of host chromosomal DNA in the final plasmid product. Involves a series of new unit operations and devices for cell resuspension, cell lysis, and neutralization.

Abstract: A scalable alkaline lysis process, including procedures and devices for the isolation of large quantities (grams and kilograms) of plasmid DNA from recombinant E. coli cell. Effective, contgrollable, and economical operation, and consistent low level of host chromosomal DNA in the final plasmid product. Involves a series of new unit operations and devices for cell resuspension, cell lysis, and nuetralization.

Abstract: Cationic lipids are provided which are useful in the preparation of liposomes and other lipid vesicle carriers. The lipids of the invention are particularly useful as carriers of nucleic acids and other negatively charged substances, for delivery to cells.

Abstract: The present invention provides mutant proteins of steroid hormone receptors. These mutant proteins are useful in methods of distinguishing a steroid hormone receptor antagonist from a steroid hormone receptor agonist. The present invention also provides plasmids containing mutated steroid hormone receptor proteins and cells transfected with those plasmids. In addition, the present invention provides methods for determining whether a compound is a steroid hormone receptor antagonist or agonist. Also, the present invention provides methods of determining endogenous ligands for steroid hormone receptors. The invention further provides a molecular switch protein for regulating expression in gene therapy.

Abstract: A nucleic acid formulation for use in gene delivery comprising a nucleic acid and an anionic polymer is disclosed. Examples of the anionic polymer includes aniionic amino acid polymer or poly-amino acid (such as poly-L-glutamic acid, poly-D-glutamic acid, poly-L-aspartic acid, poly-D-aspartic acid), poly-acrylic acid, polynucleotides, poly galacturonic acid, and poly vinyl sulfate.

Abstract: Methods are provided for producing highly purified compositions of nucleic acids by using tangential flow ultrafiltration. A scaleable process for producing pharmaceutical grade plasmid DNA, useful for gene therapy, is provided, which is efficient and avoids the use of toxic organic chemicals.

Abstract: The present invention, as noted above, relates generally to the incorporation of plasmid into a conventional dosage form, and more particularly to the production of a single-vial, homogenized, plasmid/polymer complex with desirable physical characteristics. Methods of making, storing and using such a complex are also provided and described in detail below. Such products and methods will provide more convenient and cost-effective complexes, which will be protected against chemical degradation and/or physical aggregation of its components and will provide for relative ease of administration. Thus, the present invention provides a more efficient complex for plasmid delivery and a method of incorporation of that plasmid into a conventional dosage form.

Abstract: Compositions and methods for administering nucleic acid compositions in vitro to cells in culture or in vivo to an organism whereby the uptake of nucleic acids is enhanced are provided. Various compositions, including those incorporating protective, interactive, non-condensing compounds, are utilized to protect and administered nucleic acid formulation, thereby prolonging the localized bioavailability of the administered nucleic acid and enhancing expression from the nucleic acid.

Abstract: This invention provides polypeptides having anti-angiogenic activity and nucleic acids that encode these polypeptides. The anti-angiogenic polypeptides include at least kringles 1-3 of plasminogen. The invention also provides methods of using the polypeptides and nucleic acids for inhibiting angiogenesis and other conditions characterized by undesirable endothelial cell proliferation.

Abstract: The present invention provides improved methods for detection of recombinant proteins. The fusion proteins of the invention comprise a capture tag sequence, a detection tag sequence, and polypeptide sequence of interest.

Abstract: Vectors which establish controlled expression of recombinant GHRH genes within tissues at certain levels. The vector includes a 5′ flanking region which includes necessary sequences for expression of a nucleic acid cassette, a 3′ flanking region including a 3′UTR and/or 3′NCR, and a linker which connects the 5′ flanking region to a nucleic acid sequence. The linker has a position for inserting a nucleic acid cassette. The linker does not contain the coding sequence of a gene that the linker is naturally associated with. The 3′ flanking region is 3′ to the position for inserting the nucleic acid cassette.

Abstract: The present invention provides mutated proteins of steroid hormone receptors. These mutated proteins are useful as gene medicines. In particular, these mutated proteins are useful for regulating expression of genes in gene therapy. In addition, the present invention provides plasmids encoding for the desired mutated steroid hormone receptor proteins, as well as cells transfected with those plasmids.

Abstract: The invention concerns making and evaluating synthetic regulatory regions for controlling gene expression. The invention features a method for identifying transcription factor binding sites and a method for evaluating the regulatory functions of synthetic regulatory regions.

Abstract: A novel method is provided for delivering nucleic acid molecules to the cells of an organism by pulse voltage delivery. The method involves the combination of formulated nucleic acid molecules with devices for injecting the molecules by pulse voltage or an electrical field. Disclosed are compositions and methods for enhancing the administration to and uptake of nucleic acids in a mammal. The methods disclosed provide an increased transfection and/or gene delivery efficiency by enhancing the uptake of formulated nucleic acid molecules by applying an electrical field which destabilizes the cellular membrane thereby opening pores or passageways which allow extracellular material to be introduced to the cell. Also disclosed are examples which demonstrate that the combination of formulated nucleic acid molecules and pulse voltage injection methods results in immune responses which are superior to those obtained by conventional means of delivery.

Abstract: This invention herein describes pharmaceutical compositions and methods for targeted delivery of functional genes into cells and tissues in vivo. The invention discloses DNA:lipid complexes, methods of making such complexes and methods of using such complexes for facilitating the targeted delivery and entry of recombinant expression constructs into cells and tissues in vivo, and particularly delivery of such recombinant expression constructs by intravenous, intraperitoneal or direct injection.

Abstract: A scalable alkaline lysis process, including procedures and devices for the isolation of large quantities (grams and kilograms) of plasmid DNA from recombinant E. coli cells. Effective, controllable, and economical operation, and consistent low level of host chromosomal DNA in the final plasmid product. Involves a series of new unit operations and devices for cell resuspension, cell lysis, and neutralization.

Abstract: Methods are provided for the preparation of transfection complexes of polynucleotides and polycations, especially cationic lipids, suitable for delivering polynucleotides to cells. In particular, methods are provided for preparing transfection complexes using a reduced-volume, dual-feed process. Complexes are formed upon the collision of two feed stream, containing polynucleotides and polycation, respectively, under conditions resulting in turbulent mixing conditions in minimal volume, and removal of transfection complexes under laminar flow conditions. Alternatively, the components are mixed in a static mixer. The process is easily scaleable and highly reproducible.

Abstract: Compositions and methods are provided for gene delivery to the respiratory tract. In particular, compositions comprising a nebulized nucleic acid/stabilizing agent complex, and methods employing such complexes for pulmonary gene delivery, are provided. Such complexes are preferably sonic nebulized.