Abstract: Thermostable enzymes are subjected to mutagenesis to produce a thermophilic enzyme which is stable at thermophilic temperature and which has increased activities at least two-fold higher than the activity of the wild-type thermostable enzyme at lower temperatures, which are temperatures of 50° C. or lower.

Abstract: This invention provides methods of obtaining novel polynucleotides and encoded polypeptides by use of non-stochastic methods of directed evolution (DirectEvolution™). These methods include non-stochastic polynucleotide site-saturation mutagenesis (Gene Site Saturation Mutagenesis™) and non-stochastic polynucleotide reassembly (GeneReassembly™). Through use of the claimed methods, genetic vaccines, enzymes, and other desirable molecules can be evolved towards desirable properties. For example, vaccine vectors can be obtained that exhibit increased efficacy for use as genetic vaccines. Vectors obtained by using the methods can have, for example, enhanced antigen expression, increased uptake into a cell, increased stability in a cell, ability to tailor an immune response, and the like. This invention provides methods of obtaining novel enzymes that have optimized physical &/or biological properties.

Abstract: This invention provides methods of obtaining novel polynucleotides and encoded polypeptides by the use of non-stochastic methods of directed evolution (DirectEvolution™). A particular advantage of end-selection-based methods is the ability to recover full-length polynucleotides from a library of progeny molecules generated by mutagenesis methods. These methods include non-stochastic polynucleotide site-saturation mutagenesis (Gene Site Saturation Mutagenesis™) and non-stochastic polynucleotide reassembly (GeneReassembly™). This invention provides methods of obtaining novel enzymes that have optimized physical &/or biological properties. Through use of the claimed methods, genetic vaccines, enzymes, small molecules, and other desirable molecules can be evolved towards desirable properties. For example, vaccine vectors can be obtained that exhibit increased efficacy for use as genetic vaccines.

Abstract: A safe gene transfer system is provided based on simian immunodeficiency virus (SIV). A packaging construct is provided in which the SIV packaging sequence is removed, and, in one instance, the gag, pol and other accessory genes are expressed under the control of CMV promoter. SIV transfer constructs are provided in which cis-acting sequences required for viral replication (R, tRNA primer binding site, encapsidation site, RRE sequence and 3′ LTR), with or without a marker gene, are described. Stable packaging cell lines are provided that were prepared by transfecting 293(T) cells with an SIV packaging construct. High titer virus (>107 transducing units/ml) can be produced upon transient transfection with SIV transfer vectors. The safety of the described SIV gene transfer vectors was further tested in a primate model. Rhesus monkeys that received multiple inoculations of high titer (107) retroviral vectors did not develop any signs of side effects.

Abstract: A directed evolution process comprising novel methods for generating improved progeny molecules having desirable properties, including, for example, a method for rapid and facilitated production from a parental polynucleotide template, of a set of mutagenized progeny polynucleotides wherein at least one codon encoding each of the 20 naturally encoded amino acids is represented at each original codon position. This method, termed site-saturation mutagenesis, or simply saturation mutagenesis, is preferably based on the use of the degenerate N,N,G/T sequence. Also, a method of producing from a parental polypeptide template, a set of mutagenized progeny polypeptides wherein each of the 20 naturally encoded amino acids is represented at each original amino acid position.

Abstract: A mutant virus is provided which contains a mutation at a phosphorylation site in one or more of the proteins of the virus, which mutation causes the virus to be attenuated, and therefore, an improved vaccine composition can be produced therewith. The invention also relates to vaccine compositions which contain the mutant virus, as well as to methods of inducing an immune response, and of protecting mammals from infection by rabies virus. Also included in the invention are methods of producing the mutant virus and mutant viral proteins, including producing the mutant virus in a host cell which produces or even overproduces a wild-type counterpart of the mutant viral protein, which complements the other viral proteins such that production of the mutant viral particle is optimized.

Abstract: The invention concerns constructs for the expression of a protein comprising at least a modified FV envelope protein, the protein so obtained as well as the complementation cell line permitting the production of pseudotyped viral particle. It also concerns pharmaceutical composition comprising said particles and a method for treating a disease.

Abstract: The present invention relates to a method and an apparatus for separating substances from or in biological materials.

Abstract: The invention provides methods, reagents and devices for analysis of receptor ligand interactions, determining the profile of receptor expression in cells and cell populations, and diagnostic and drug screening methods. The invention makes use of immobilized tethered ligand fusion proteins having a ligand domain, a stalk domain, and optionally an immobilization domain.

Abstract: Synthetic DNA molecules encoding HIV gag and modifications of HIV gag are provided. The codons of the synthetic molecules are codons preferred by the projected host cell. The synthetic molecules may be used as a polynucleotide vaccine which provides effective immunoprophylaxis against HIV infection through stimulation of neutralizing antibody and cell-mediated immunity.

Abstract: The present invention provides two new HIV/SIV translocation promoting agents, Bonzo and BOB. The present invention also provides the amino acid and DNA sequences of human, African green monkey, and pigtail macaque of the receptor protein Bonzo. Mammalian cells transfected with Bonzo and/or BOB and human CD4 as well as antibodies to the receptor Bonzo are also included. Furthermore, a method of identifying other such translocation promoting agents is also disclosed. Diagnostic and therapeutic uses of the translocation promoting agents of the present invention are also provided. Furthermore, the present invention provides methods of identifying agents that can modulate the expression and/or function of Bonzo/STRL33. Such agents can be used to either treat inflamation, or alternatively, to enhance the immune response.

Abstract: A directed evolution process comprising novel methods for generating improved progeny molecules having desirable properties, including, for example, a method for rapid and facilitated production from a parental polynucleotide template, of a set of mutagenized progeny polynucleotides wherein at least one codon encoding each of the 20 naturally encoded amino acids is represented at each original codon position. This method, termed site-saturation mutagenesis, or simply saturation mutagenesis, is preferably based on the use of the degenerate N,N,G/T sequence. Also, a method of producing from a parental polypeptide template, a set of mutagenized progeny polypeptides wherein each of the 20 naturally encoded amino acids is represented at each original amino acid position.

Abstract: Liposomal conjugates having a clinically useful delivery vehicle linked to a biologically active species which acts to increase vascular permeability and expand blood volume at or in proximity to the tumor site are disclosed. The vehicle-linked species may be, for example, a vasoactive agent, a substance that recruits or amplifies a vasoactive species, a drug, or a pharmaceutical compound. Suitable biological species comprises peptides, lipids, carbohydrates, or their derivatives. Chemical or recombinant DNA methods suitable for linking the species to the vehicles are indicated. A therapy is disclosed which comprises administering the vasoactive conjugate and delivering a diagnostic agent or a therapeutic agent at an optimal time thereafter, when tumor vasculature is maximally affected.

Abstract: The invention belongs to the field of biologically active peptides specifically stimulating antiviral, antimicrobial and antitumor activity of the human and animal immune system.

Abstract: Activated lymphocytes derived from cord blood are excellently effective for preventing and treating various types of tumors and various types of infection. With interleukin 2 and/or anti-CD3 antibody, the lymphocytes derived from the cord blood is prepared by segregating lymphocytes from the cord blood and proliferating the segregated lymphocytes directly in vitro or segregating monocytes from cord blood and proliferating the monocytes in vitro. Also, the cord blood-derived activated lymphocytes can be effectively used for preventing recurrence of the diseases and promoting the take of stem cells or other organs.

Abstract: The invention relates to novel recombinant glycoproteins that act as messenger substances, signaling substances, promoters, stimulators and initiators in a multitude of ways in the animal, especially human, circulation system.

Abstract: The present invention provides truncated HCV E2 polypepides. The invention HCV E2 polypeptides lack the HVR1 region that provides immune protection against HCV. The present invention also provides immunogenic compositions of such polypeptides and the methods of use thereof.

Abstract: A recombinant vector comprising a polynucleotide containing a cis-acting central initiation region (cPPT) and a cis-acting termination region (CTS) is disclosed. These regions are of retroviral or retroviral-like origin. The vector further comprises a defined nucleotide sequence (transgene or sequence of interest) and regulatory signals for reverse transcription, expression, and packaging, wherein the regulatory signals are of retroviral or retroviral-like origin. In some embodiments the vector is incorporated into compositions for therapeutic purposes. In other embodiments the vector is incorporated into immunogenic compositions, The vector is also useful for methods such as ex vivo transfection or ex vivo transduction of non-mitotic differentiated cells.

Abstract: Methods are provided for treatment of eye disorders and injury, including methods for treatment of reduced flow of blood or other nutrients to retinal tissue and/or optic nerve, methods for treatment of retinal ischemia and trauma and methods for treatment for optic nerve injury/damage.

Abstract: This invention relates to improved methods for producing nonsegmented, negative-sense, single-stranded RNA viruses of the Order designated Mononegavirales virus, including embodiments relating to methods of producing such viruses as attenuated and/or infectious viruses, such as Measles virus (MV) and respiratory syncytial virus (RSV).