Applied Immunesciences, Inc.

Rhone-poulenc Rorer Pharmaceuticals Inc.

Aventis Pharmaceuticals Inc.

Thomas B Okarma

Jane S Lebkowski

Maureen A McNally

John Blankenship

Abraham T Lin

Mohammad A Elkalay

Brian R Clark

Chin-Hai Chang

L Bernard Lerch

David A Okrongly

Donald Lamons

Young M Lee

Kevin R Leiby

Devices, processes and compositions are provided for effective separation of cells from a mixture of cells, where depletion or positive selection may be employed to provide a cellular population of interest. Of particular utility is the separation of cells from peripheral blood mononuclear cells, where members of the lymphoid or myeloid lineages may be isolated and used for research, diagnosis or therapy. Also of interest are cellular separation from bone marrow, tumor suspensions or lymphoid tissue suspensions, where cells can be isolated and used for a variety of purposes. The separated cells may be homogeneous, free of exogenous biologicals, viable, capable of replication and exhibit their full complement of biological activities. Multiple phenotypes can be captured simultaneously. Captured cells can be specifically activated with cytokines and antigens to provide cells which are MHC restricted and have antigen-specific effector functions.

Device and process for cell capture and recovery

Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of viral elements produced by conventional methods.

Recombinant adeno-associated virus vectors

This invention provides a composition, device and method for the removal of selected factors, such as cytokines or pharmaceuticals, from a substance such as whole blood or plasma. Advantageously, the invention provides for the treatment or prevention of septic shock syndrome or other conditions evidenced by the presence of cytokines in a patient by contacting the patient's whole blood with a composition comprising silica and a surface treatment material, such as heparin, but preferably human serum albumin (HSA). The treatment lowers the cytokine concentration of the blood. Pharmaceuticals can be removed from an individual's whole blood or plasma, such as for use in treating drug overdosage.

Removal of selected factors from whole blood or its components

Recombinant adeno-associated virus vector packaging cells and methods

Method for producing recombinant adeno-associated virus vectors

Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The AAV rep and cap genes are combined in a single recombinant adenovirus vector, i.e., a rep/cap adenovirus vector which combines in a single vector all complementing functions required for rAAV production. The rep/cap vectors of the present invention comprise: genes encoding helper functions, preferably from adenovirus, sufficient to permit AAV replication; and AAV cap and rep genes. In adenovirus rep/cap vectors the rep and cap genes may, for example, replace the adenovirus E3 gene or the adenovirus E1a and E1b genes.

This invention provides a composition, device and method for the treatment or prevention of septic shock syndrome or other conditions evidenced by the presence of cytokines in a patient by contacting the patient's whole blood with a composition comprising silica and a surface treatment material, such as heparin, but preferably human serum albumin (HSA). The treatment lowers the cytokine concentration of the blood.

Removal of selected factors from whole blood or its components; and

Simplified methods to produce recombinant adeno-associated virus (rAAV) vectors are described. The methods involve the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr virus (oriP), and a rAAV genome. The chimeric plasmids themselves are also a part of the present invention. These plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of rAAV upon addition of wild-type, adeno-associated virus helper functions. Vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit the attributes of vital elements produced by conventional methods.

Production of recombinant adeno-associated virus vectors

Peptide fragments of conglutinin are provided for use in binding to complementary ligands. Particularly, an N-proximal region is provided having a hypervariable region with a collagen type structure for binding to complementary molecules, and a C-proximal region which provides for lectin binding activity.

C-terminal conglutinin fragments coupled to solid support

A method and composition which permits sterilization of surfaces coupled with biologically active moieties by ionizing radiation is described. The protecting composition contains a surface-stabilizing agent which adheres to the surface and has a molecular weight.gtoreq.5 kd, and an oxygen radical scavenger which is preferably a di- or polysaccharide or reduced form thereof. In the method of the invention, a surface which is coupled to a biologically active agent is protected with the invention composition, dried to a moisture content of less than 1%, and then sterilized by ionizing radiation under standard conditions. The sterilized surfaces of the invention are particularly useful in the production of medical devices intended for extracorporeal use, particularly in cell-separation techniques.

Stabilization of sterilized surfaces for research and medical use

A simplified method to produce recombinant adeno-associated virus (AAV) vectors is described. The procedure involves the use of chimeric plasmids which incorporate the Epstein Barr nuclear antigen (EBNA) gene, the latent origin of replication of Epstein Barr Virus (oriP), and a recombinant AAV genome. The chimeric plasmids themselves are also a part of the present invention. These EBV/AAV plasmids are maintained as multicopy extra-chromosomal elements in cells, such as human 293 cells. Permanent cell lines carrying these EBV/AAV plasmids are induced to produce large amounts of recombinant AAV virus upon addition of wild-type, adeno-associated virus helper functions. Recombinant AAV vectors produced in this manner are capable of transducing exogenous genes into other human cell lines and exhibit all of the attributes of viral elements produced by conventional methods.

Compositions and devices are provided for the specific removal of components of plasma in efficient and economical ways. The devices provide for a tortuous path of the plasma through a high surface material to which is bound a binding compound for removal of the fluid component. The devices find particular application with plasma, in diagnosis, therapy, and for production of specific physiologically active materials.

Device for plasma modification--composition and remodeling

Blood derived fluids are subjected to modification resulting in the production of anaphylatoxins. The anaphylatoxins may then be removed by passing the modified blood through silicic acid particles in an amount sufficient to substantially reduce the anaphylatoxins, while still retaining the other blood components and without affecting adversely the use of the blood for the patient.

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Palo Alto, CA, United States of America

Thomas B Okarma