Fusion partner cells and uses thereof
AssigneeWhitehead Institute for Biomedical Research
InventorsScott K. Dessain, Robert A. Weinberg
Life of this patent
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Abstract
The present invention provides in one aspect novel fusion partner cells that ectopically express one or more genes that alter the phenotype of a hybrid cell made from a fusion of the fusion partner cell and a fusion cell, hybrid cell lines produced using the fusion partner cells. The invention in another aspect provides antibodies produced by certain hybrid cell lines, and compositions containing one or a combination of such antibodies or antigen-binding fragments thereof. The invention also provides in another aspect methods of using the antibodies or antigen-binding fragments thereof for diagnosis and treatment of diseases characterized by the antigens specifically bound by the antibodies.
Claims157
A fusion partner cell comprising at least two ectopically expressed nucleic acid molecules, wherein each of the ectopically expressed nucleic acid molecules encodes a polypeptide that when expressed in the hybrid cell, alters the phenotype of the hybrid cell.
A fusion partner cell comprising at least one ectopically expressed nucleic acid molecule that regulates the expression of at least one polypeptide that when expressed in the hybrid cell, alters the phenotype of the hybrid cell.
A fusion partner cell comprising a soluble or membrane bound growth factor selected from the group consisting of IL-6 and IL-11 and at least one ectopically expressed nucleic acid molecule that encodes at least one polypeptide that when expressed in the hybrid cell, alters the phenotype of the hybrid cell.
A fusion partner cell comprising an ectopically expressed nucleic acid molecule that encodes a polypeptide that inhibits tumor suppressor activity.
A fusion partner cell comprising an ectopically expressed nucleic acid molecule that encodes a growth promoting polypeptide, wherein the nucleic acid is derived from a different species than the cell.
A human fusion partner cell comprising an ectopically expressed nucleic acid molecule that encodes a growth promoting polypeptide, wherein the nucleic acid is derived from a human.
A method for making immunoglobulin-secreting hybrid cells comprising
- fusing B lymphocytes to a fusion partner cell comprising at least one ectopically expressed nucleic acid molecule, wherein the ectopically expressed nucleic acid molecule encodes a polypeptide that when expressed in the hybrid cell, alters the phenotype of the hybrid cell, to form hybrid cells, thereby producing immunoglobulin secreting hybrid cells.
A method for making immunoglobulin-secreting hybrid cells comprising
- fusing B lymphocytes to a fusion partner cell to form immunoglobulin secreting hybrid cells, and
- ectopically expressing at least one nucleic acid molecule that encodes a polypeptide that when expressed in the hybrid cell, alters the phenotype of the hybrid cell.
A method for making immunoglobulin molecules comprising
- fusing B lymphocytes to the fusion partner cells of claim I to form hybrid cells, and
- maintaining resulting hybrid cells under conditions appropriate for production of immunoglobulin molecules by hybrid cells, whereby immunoglobulin molecules are produced by hybrid cells.
A method for cloning immunoglobulin-encoding nucleotide sequences, comprising
- preparing a library of human hybridoma cells,
- selecting from the library one or more immunoglobulin-secreting cells of interest,
- isolating immunoglobulin-encoding nucleotide sequences from the selected immunoglobulin-secreting cells.
A method for producing an antibody with a desired specificity, comprising:
- preparing a library of hybridoma pools,
- performing limiting dilution on the hybridoma pools,
- analyzing antibody produced by the hybridoma pools to identify a putative antibody with a desired specificity,
- cloning immunoglobulin genes from hybridoma pools that produce the putative antibody, and
- expressing the immunoglobulin genes in a host cell to produce an antibody with desired specificity.
A method of identifying an antibody developed in a human in response to exposure of the immune system of the human to an antigen, the method comprising the steps of:
- (a) generating fused cells by mixing together under fusing conditions:
- human B cells with culturable fusion partner cells;
- (b) detecting a subset of surviving fused cells which express an antibody that selectively binds the antigen;
- (c) isolating nucleotide sequences encoding at least the CDRs of the antibody from the subset of surviving fused cells;
- (d) transfecting nucleotide sequences isolated in (c) into a culturable cell line to produce a plurality of culturable cells expressing antibodies comprising the CDRs;
- and
- (e) screening culturable cells produced in (d) to detect an antibody comprising the CDRs which binds to the antigen, thereby identifying an antibody.
A hybridoma comprising a fusion partner cell comprising at least one ectopically expressed nucleic acid molecule, wherein each of the ectopically expressed nucleic acid molecule encodes a polypeptide that when expressed in the hybrid cell, alters the phenotype of the hybrid cell fused to a primary mammalian cell, wherein the primary mammalian cell and the fusion partner cell are derived from different species.
Dependencies are inferred from the claim text.