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OwnerLuigi Grasso, J. Bradford Kline +2inventors
InventorsLuigi Grasso, J. Bradford Kline, Nicholas C. Nicolaides, Philip M. Sass
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The use of mismatch repair (MMR) defective antibody producer cells offers a method to generate subclone variants with elevated protein production such as antibodies. Using MMR defective cells and animals, new cell lines and animal varieties with novel and useful properties such as enhanced protein production can be generated more efficiently than by relying on the natural rate of mutation. These methods are useful for generating genetic diversity within host cells to alter endogenous genes that can yield increased titer levels of protein production. By employing this method, two genes were discovered whose suppressed expression is associated with enhanced antibody production. Suppressed expression of these genes by a variety of methods leads to increased antibody production for manufacturing as well as strategies for modulating antibody production in immunological disorders. Moreover, the suppression of these two genes in host cells can be useful for generating universal high titer protein production lines.
A method for identifying genes responsible for high titer antibody production comprising:
A homogeneous culture of high titer antibody producing cells produced by a method comprising the steps of:
A method for producing a high titer antibody producing cell comprising the step of modulating the expression of at least one gene involved in antibody production wherein said genes comprise alpha1-anti-trypsin and endothelial monocyte-activating polypeptide I.
A method of modulating antibody production of cells comprising contacting said cells with protease inhibitors to decrease antibody production from antibody producer cells.
The method of claim 33 where the inhibitor comprises pharmacological amounts of natural protease substrates.
A method for selecting cells for high titer antibody production whereby growth medium of cells is analyzed for alpha-1-antitrypsin, where low levels are associated with high antibody titers.
A method for selecting for cells for high titer antibody production whereby growth medium of cells is analyzed for endothelial monocyte-activating polypeptide I, where low levels are associated with high antibody titers.
A method for suppressing antibody production associated with hyperimmunoglobulin disease production comprising contacting said cells with at least one compound that increases endothelial monocyte-activating polypeptide I expression.
A method for suppressing antibody production associated with hyperimmunoglobulin disease production comprising contacting said cells with at least one compound that increases alpha-1-antitrypsin expression.
A method for enhancing antibody production associated with hypoimmunoglobulin disease production comprising contacting said cells with at least one compound that suppresses alpha-1-anti-trypsin expression activity.
A method for enhancing antibody production associated with hypoimmunoglobulin disease production comprising contacting said cells with at least one compound that suppresses monocyte-activating polypeptide I expression activity.
A host cell for the expression of antibody molecules or fragments thereof comprising a defect in the monocyte-activating polypeptide I gene such that expression of monocyte-activating polypeptide I is inhibited.
A host cell for the expression of antibody molecules or fragments thereof comprising a defect in the alpha-1-antitrypsin gene such that expression of alpha-1-antitrypsin is inhibited.
Dependencies are inferred from the claim text.