Abstract: The present invention relates to improved human FVIII variants having at least one substitution in the A2 and/or C2 domain. The present invention also relates to their uses in the treatment of hemophilia A, particularly in patients with inhibitors.
Type:
Grant
Filed:
February 22, 2008
Date of Patent:
January 7, 2014
Assignees:
Biomethodes, Hospices Civils de Lyon
Inventors:
Didier Saboulard, Jean-Luc Plantier, Marc Delcourt, Claude Negrier, Thierry Menguy, Stephane Blesa, Sylvie Marin
Abstract: The invention concerns the field of molecular biology and more particularly that of mutagenesis. It concerns a method of high-rate directed mutagenesis, that is the formation of numerous directed mutants in reduced time and with reduced number of steps. Said method is therefore referred to as massive mutagenesis.
Abstract: The present invention pertains to a process for isolating an intact clone of one target nucleic acid fragment having a known characteristic, from a group of fragments by preparing an initial library of clones from the group of fragments using a vector containing no more than a predetermined number of known restriction sites, preferably 1–3 restriction sites, subjecting the initial library to at least 10, and preferably between 50 and 70 restriction enzymes different from those to which the vector is susceptible, to produce a group of monodigested libraries, screening the group of monodigested libraries for the target fragment to determine those restriction enzymes to which the target fragment is insensitive, and subjecting the initial library to substantially all of the restriction enzymes to which the target fragment is insensitive, to produce a multidigested library having an intact clone of the target nucleic acid fragment.