Methods of generating hybrid/chimeric cells, and uses thereof
Inventors
Kaseko, Galina • Mahaworasilpa, Tohsak L.
Assignees
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Publication Number
US-8778677-B2
Publication Date
2014-07-15
Expiration Date
Abstract
The present invention relates to hybrid cells and methods for producing hybrid cells. In particular, the invention relates to hybrid cells generated from the hybridization of at least three cells where at least two cells are derived from different lineages. The invention further relates to the use of hybrid cells for the expression of proteins useful in a range of diagnostic, prophylactic, therapeutic and/or research applications.
Core Innovation
The invention relates to a hybrid cell generated by hybridisation of a first cell derived from a common myeloid progenitor cell, and a second and a third cell derived from a common lymphoid progenitor cell. The first cell may optionally be a myelomonocytic progenitor, monocyte, macrophage, eosinophil, neutrophil, dendritic cell or basophil. The hybridisation is cross-lineage, combining myeloid and lymphoid progenitor-derived cells in a tri-hybrid configuration.
A critical exclusion is that the second cell and the third cell are not myeloma cells. The hybrid cell is therefore a myeloma-free hybridisation configuration while maintaining a hybrid lineage relationship between the myeloid-progenitor-derived component and two lymphoid-progenitor-derived components.
The document describes cross-lineage chimeric tri-hybrid cell lines and reports that established cross-lineage tri-hybrids are EBV-genome negative and mycoplasma-free. The tri-hybrid system supports protein expression, including examples such as hGM-CSF, human immunoglobulins, and hCD54, with functional characterization that verifies glycosylation- and marker-dependent features.
Claims Coverage
The claim coverage centers on one independent hybrid-cell claim and dependent refinements. The independent claim contains three inventive combination elements: a myeloid-progenitor-derived first cell, two lymphoid-progenitor-derived second and third cells, and the explicit requirement that the second and third cells are not myeloma cells.
Myeloma-free tri-hybrid cross-lineage cell
A hybrid cell generated by hybridisation of: a first cell derived from a common myeloid progenitor cell, a second cell derived from a common lymphoid progenitor cell, and a third cell derived from a common lymphoid progenitor cell, wherein the second cell and the third cell are not myeloma cells.
Overall claim coverage centers on a tri-hybrid, cross-lineage construct combining a common myeloid progenitor-derived component with two common lymphoid progenitor-derived components, while excluding myeloma cells from the lymphoid-derived components. Dependent refinements described in the provided material further constrain hybridisation methods and downstream protein expression and selection by cell type-defining CD markers.
Stated Advantages
Human-like post-translational modifications.
Stable functionality.
Enhanced expression upon co-expression of additional proteins.
Documented Applications
Protein expression for diagnostic, prophylactic, therapeutic, and research uses.
Use of the cross-lineage tri-hybrid system to support protein expression, with functional characterization verifying glycosylation- and marker-dependent features (examples include hGM-CSF, human immunoglobulins, and hCD54).
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