Abstract: This disclosure provides methods and landing pad constructs for generation of parental cell lines suitable for targeted integration. A method is provided by the parental cell line development; this is, the introduction of binding sites of BPV1 E2 protein to landing pad vectors so that expressed BPV1 E2 protein could locate the vector to transcriptionally active region in the genome. Cells with high expression level of reporter genes are selected for the next stage and will be used in the development of cell lines expressing another recombinant protein by recombination mediated cassette exchange (RMCE). Landing pad constructs include recombination target sites for site-specific recombinases, and therefore, it could be replaced with gene-of-interest expression construct containing the same set of recombination target sites. This yields the generation of producer cell lines with less effort compared to traditional cell line development by random integration.

Abstract: This disclosure provides methods and landing pad constructs for generation of parental cell lines suitable for targeted integration. A method is provided by the parental cell line development; this is, the introduction of binding sites of BPV1 E2 protein to landing pad vectors so that expressed BPV1 E2 protein could locate the vector to transcriptionally active region in the genome. Cells with high expression level of reporter genes are selected for the next stage and will be used in the development of cell lines expressing another recombinant protein by recombination mediated cassette exchange (RMCE). Landing pad constructs include recombination target sites for site-specific recombinases, and therefore, it could be replaced with gene-of-interest expression construct containing the same set of recombination target sites. This yields the generation of producer cell lines with less effort compared to traditional cell line development by random integration.

Abstract: Methods for treating high risk HPV infection by inhibiting one or more phases of HPV replication cycle with antiviral compounds are disclosed. Moreover, a mechanism for the inhibition is suggested and targets for further antiviral compounds are disclosed.

Abstract: This invention provides a method and a kit for rapid and robust development monoclonal antibodies. The method does not require hybrid technologies nor does it require single cell manipulations. The method allows elimination of cross-target antibodies.

Abstract: A U2OS-based model system using luciferase reporters to monitor the genome replication of alpha and beta HPVs is provided. A modified U2OS cell line is disclosed that expresses Firefly luciferase to measure toxicity of the screened compounds. In addition, provided are HPV18, HPV 16 and HPV5 marker genomes that express Renilla luciferase under the control of viral promoters used to measure changes in the viral copy number. This ready-to-use model system is capable of being used in high-throughput screens to identify compounds inhibiting initial amplification and stable maintenance as well as vegetative phase of various HPV subtypes.

Abstract: This disclosure provides methods and landing pad constructs for generation of parental cell lines suitable for targeted integration. A method is provided by the parental cell line development; this is, the introduction of binding sites of BPV1 E2 protein to landing pad vectors so that expressed BPV1 E2 protein could locate the vector to transcriptionally active region in the genome. Cells with high expression level of reporter genes are selected for the next stage and will be used in the development of cell lines expressing another recombinant protein by recombination mediated cassette exchange (RMCE). Landing pad constructs include recombination target sites for site-specific recombinases, and therefore, it could be replaced with gene-of-interest expression construct containing the same set of recombination target sites. This yields the generation of producer cell lines with less effort compared to traditional cell line development by random integration.

Abstract: Methods for treating high risk HPV infection by inhibiting one or more phases of HPV replication cycle with antiviral compounds are disclosed. Moreover, a mechanism for the inhibition is suggested and targets for further antiviral compounds are disclosed.

Abstract: Novel antiviral compounds for inhibiting one or more phases of HPV replication cycle are disclosed. Moreover, a mechanism for the inhibition is suggested and targets for further antiviral compounds are disclosed.

Abstract: This invention provides a method and a kit for rapid and robust development monoclonal antibodies. The method does not require hybrid technologies nor does it require single cell manipulations. The method allows elimination of cross-target antibodies.

Abstract: A U2OS-based model system using luciferase reporters to monitor the genome replication of alpha and beta HPVs is provided. A modified U2OS cell line is disclosed that expresses Firefly luciferase to measure toxicity of the screened compounds. In addition, provided are HPV18, HPV 16 and HPV5 marker genomes that express Renilla luciferase under the control of viral promoters used to measure changes in the viral copy number. This ready-to-use model system is capable of being used in high-throughput screens to identify compounds inhibiting initial amplification and stable maintenance as well as vegetative phase of various HPV subtypes.

Abstract: Novel antiviral compounds for inhibiting one or more phases of HPV replication cycle are disclosed. Moreover, a mechanism for the inhibition is suggested and targets for further antiviral compounds are disclosed.

Abstract: A method for treating an HIV disease in a subject in need of said treatment, comprising administering to the subject a therapeutically effective amount of a DNA vaccine comprising an expression vector and a pharmaceutically acceptable excipient, where the expression vector comprises: (a) a heterologous promoter operatively linked to a DNA sequence encoding a nuclear-anchoring protein, where the nuclear-anchoring protein comprises: (i) a DNA binding domain which binds to a specific DNA binding sequence, and (ii) a functional domain of the Bovine Papilloma Virus Type 1 E2 protein, where the functional domain binds to a nuclear component; (b) a multimerized DNA sequence that forms a binding site for the nuclear anchoring protein; and (c) at least one expression cassette comprising a DNA sequence encoding a protein or peptide that stimulates an immune response specific to the protein or peptide; where the expression vector lacks an origin of replication functional in mammalian cells.

Abstract: The present invention relates to an alphaviral replicase, especially Semliki Forest Virus replicase, or an expression vector encoding an alphaviral replicase, said alphaviral replicase comprising RNA dependent RNA polymerase activity, for use as an immune system modulating adjuvant. The alphaviral replicase may be used in the combination with a vaccine providing an adjuvant function therein, which when present therein will generate an additional boost to the immune response in the subject to whom this combination is administered as compared to when the vaccine alone is administered to a subject in need thereof. The aim of the present invention is to provide an efficient and easy to administer, species-independent adjuvant which will provide advantages to the adjuvants used together with vaccines today.

Abstract: A method for treating an HIV disease in a subject in need of said treatment, comprising administering to the subject a therapeutically effective amount of a DNA vaccine comprising an expression vector and a pharmaceutically acceptable excipient, where the expression vector comprises: (a) a heterologous promoter operatively linked to a DNA sequence encoding a nuclear-anchoring protein, where the nuclear-anchoring protein comprises: (i) a DNA binding domain which binds to a specific DNA binding sequence, and (ii) a functional domain of the Bovine Papilloma Virus Type 1 E2 protein, where the functional domain binds to a nuclear component; (b) a multimerized DNA sequence that forms a binding site for the nuclear anchoring protein; and (c) at least one expression cassette comprising a DNA sequence encoding a protein or peptide that stimulates an immune response specific to the protein or peptide; where the expression vector lacks an origin of replication functional in mammalian cells.

Abstract: The present invention relates to an alphaviral replicase, especially Semliki Forest Virus replicase, or an expression vector encoding an alphaviral replicase, said alphaviral replicase comprising RNA dependent RNA polymerase activity, for use as an immune system modulating adjuvant. The alphaviral replicase may be used in the combination with a vaccine providing an adjuvant function therein, which when present therein will generate an additional boost to the immune response in the subject to whom this combination is administered as compared to when the vaccine alone is administered to a subject in need thereof. The aim of the present invention is to provide an efficient and easy to administer, species-independent adjuvant which will provide advantages to the adjuvants used together with vaccines today.

Abstract: A selection system free of antibiotic resistance genes, which is based on the use of an araD gene as a selection marker carried on a vector which is inserted in a bacterial strain deficient of the araD gene. The araD gene from E. coli encodes the L-ribulose-5-phosphate-4-epimerase. A method of selecting the cells transformed with a plasmid, which contains the araD gene. The non-antibiotic selection marker makes the system suitable for producing therapeutics. The araD gene is not essential for growth of the host but manipulation of it affects the growth under certain selective conditions. Deletion of araD leads to accumulation of substance which is toxic to the host but not to humans. The araD gene is relatively small and therefore a small plasmid may be constructed, which requires less energy for replication, and leads to increased growth rate and yield.

Abstract: The present invention relates to novel vectors, to DNA vaccines and gene therapeutics containing said vectors, to methods for the preparation of the vectors and DNA vaccines and gene therapeutics, and to therapeutic uses of said vectors. More specifically, the present invention relates to novel vectors comprising an expression cassette of a gene of a nuclear-anchoring protein, which contains a DNA binding domain capable of binding to a specific DNA sequence and a functional domain capable of binding to a nuclear component and a multimerized DNA sequence forming a binding site for the nuclear-anchoring protein, and optionally an expression cassette of a gene, genes or a DNA sequence or DNA sequences of interest. The present invention further relates to DNA vaccines and gene therapeutics containing the novel vectors, to methods for the preparation of the novel vectors and the DNA vaccines and gene therapeutics.