Genzyme Corporation

Other

Alan E Smith

Richard J Gregory

Donna Armentano

Seng H Cheng

Larry A Couture

Catherine E O'Riordan

Henry Hoppe, Iv

Shaona Lee Fang

John Marshall

Harry M Meade

Catherine R O'Riordan

Paul Ditullio

Karl M Ebert

Kathleen M Hehir

Gillian E Francis

Christina Delgado

Sucharita Paul

Amy E Erickson

Amy L (Ericson) Helgerson

Amy L Helgerson

Vincent Parkes

The present invention relates to the purification of large scale quantities of active (infectious) adenovirus and AAV, especially for use in therapeutic applications. In particular, the invention provides improved methods for contacting such viruses with suitable chromatographic materials in a fashion such that any damage to the virus, particularly to surface components thereof, resulting from contact with such chromatographic materials is minimized or eliminated. The result is the ability to rapidly and efficiently purify commercial level quantities of active (infectious) virus suitable for use in therapeutic applications, e.g. gene transfer/therapy procedures.

Purification of adenovirus and AAV

Gene Therapy vectors, which are especially useful for cystic fibrosis, and methods for using the vectors are disclosed.

Adenovirus vectors for gene therapy

Disclosed are full length isolated DNAs encoding cystic fibrosis transmembrane conductance regulator (CFTR) protein and a variety of mutants thereof. Also disclosed are antibodies specific for various CFTR domains and methods for their production. Expression of CFTR from cells transformed with these CFTR genes or cDNAs demonstrate surprising CFTR intracellular distributions and results thereby providing for new diagnostic and therapeutic procedures.

DNA molecules stabilized for propagation in bacterial cells that encode cystic fibrosis transmembrane conductance regulator

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.

Production of membrane proteins in the milk of transgenic mammals

Viruses are modified by coupling a polymer such as polyethylene glycol to obtain polymer-modified viruses that can exhibit reduced antigenicity while retaining infectivity, and which may exhibit increased circulation time in vivo. The polymer may be directly covalently attached or indirectly covalently attached via an intermediate coupling moiety to the virus. The polymer may also be indirectly noncovalently attached to the virus via a ligand such as an antibody having specificity for a viral surface component. To prepare the polymer-modified virus, the polymer is activated and coupled to the virus. A preferred activated polymer is tresyl-monomethoxypolyethylene glycol having an average molecular weight of about 5000 daltons. The polymer-modified viruses have utility for therapeutic and diagnostic in vivo applications, and may be used to introduce a transgene into a target cell by infection, or be administered to a subject having a tumor where the polymer-modified virus localizes to the tumor.

Tresyl-monomethoxypolyethylene glycol-modified viruses having viral infectivity

Production of membrane proteins in the milk of transgenic nonhuman mammals

Chromatographic purification of adeno-associated virus (AAV)

Gene therapy for cystic fibrosis

Adenoviral vectors which contain deletions of the early regions and/or late genes provide efficient delivery and expression of foreign nucleic acids of interest to patients. These vectors have a particular use in the treatment of cystic fibrosis patients. Furthermore, PAV vectors provide for a second generation of adenoviral vectors that contain the 5' ITR's, the packaging signal and the E1A enhancer. Other adenoviral vectors contain a deletion of the E1 region or a deletion of E4 but retain orf3 or orf6, and can either retain or delete the E3 region.

Adenoviral vectors for gene therapy containing deletions in the

A chimeric adenoviral vector is provided that comprises nucleotide sequence of a first adenovirus, wherein at least one gene of said first adenovirus encoding a protein that facilitates binding of said vector to a target mammalian cell, or internalization thereof within said cell, is replaced by the corresponding gene from a second adenovirus belonging to subgroup D, said vector further comprising a transgene operably linked to a eucaryotic promoter to allow for expression therefrom in a mammalian cell. Additionally, a method of delivering transgenes to target mammalian cells, particularly airway epithelial cells, is provided.

Chimeric adenoviral vectors

A threaded jar funnel for easy pouring of material into a jar, without leakage or spillage, has an upper part, with upper and lower conical segments, and a lower cylindrical part having threads on the inside. The upper conical segment of the upper part has a small angle to the vertical and the lower conical segment has a larger angle to the vertical than the upper conical segment. The upper conical segment merges into the lower conical segment to provide an aesthetically pleasing appearance, larger funnel capacity and smoother flow of contents into a jar coupled to the funnel. The lower cylindrical threaded part has a rim, which provides a tight seal when the funnel is coupled to a jar.

Threaded jar funnel

Methods for treating cystic fibrosis are described. The methods involve the administration of a protein enhancing agent, differentiating agent and/or carboxy-compound to a subject afflicted with cystic fibrosis such that mutant cystic fibrosis transmembrane regulator protein present within cystic fibrosis-associated cells becomes functional. Other aspects described include therapeutic compositions and packaged drugs.

Methods and therapeutic compositions for treating cystic fibrosis

A DNA comprising at least one inactivated hormone responsive element and a nucleic acid sequence encoding a membrane-associated protein is described. Therapeutic compositions and cells including the DNA are also described. Other aspects of the invention include methods of treating subjects having cystic fibrosis which include administering an effective amount of the DNA to subjects having cystic fibrosis such that functional cystic fibrosis transmembrane conductance regulator is produced by the subject at a level which is not detrimental to the subject. The present invention also pertains to a method of introducing the DNA into a cell such that the membrane-associated protein is produced at a level which is not detrimental to the cell and cells produced by this method. Still other aspects of the invention include a method of assaying DNA for the presence or absence of a hormone responsive element in a species in which the hormone responsive element is functional and a method of selectively breeding female transgenic mammals which produce a protein of interest.

Growing community of inventors

Dover, MA, United States of America

Alan E Smith