Multicellular living organisms and unmodified parts thereof and – Method of using a transgenic nonhuman animal to manufacture... – The protein is isolated or extracted from blood or serum
Patent
1995-03-29
2000-10-10
Hauda, Karen M.
Multicellular living organisms and unmodified parts thereof and
Method of using a transgenic nonhuman animal to manufacture...
The protein is isolated or extracted from blood or serum
800 13, 800 18, 800 4, 435463, 435 691, 435326, C12N 1585, C12N 1500, A01K 6700
Patent
active
06130364&
ABSTRACT:
A method to produce a cell expressing an antibody from a genomic sequence of the cell comprising a modified immunoglobulin locus using Cre-mediated site-specific recombination is disclosed. The method involves first transfecting an antibody-producing cell with a homology-targeting vector comprising a lox site and a targeting sequence homologous to a first DNA sequence adjacent to the region of the immunoglobulin loci of the genomic sequence which is to be converted to a modified region, so the first lox site is inserted into the genomic sequence via site-specific homologous recombination. Then the cell is transfected with a lox-targeting vector comprising a second lox site suitable for Cre-mediated recombination with the integrated lox site and a modifying sequence to convert the region of the immunoglobulin loci to the modified region. This conversion is performed by interacting the lox sites with Cre in vivo, so that the modifying sequence inserts into the genomic sequence via Cre-mediated site-specific recombination of the lox sites. Also disclosed are a form of the method used to produce a cell expressing a modified antibody molecule using Cre-mediated site-specific recombination, and antibody-producing cells obtainable by the disclosed methods. Class-switching modifications of human antibodies produced in murine hybridoma cells are exemplified.
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Jakobovits Aya
Zsebo Krisztina M.
Abgenix, Inc.
Beckerleg AnneMarie S.
Borden Paula A.
Francis Carol L.
Hauda Karen M.
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