Method of intracellular binding of target molecules
AssigneeDana-Farber Cancer Institute, Inc.
InventorsWayne A. Marasco, William A. Haseltine
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Abstract
The present invention relates to a method by which one can target an undesired target molecule or target antigen, preferably a protein. The method comprises the intracellular expression of an antibody capable of binding to the target. A DNA sequence is delivered to a cell, the DNA sequence contains a sufficient number of nucleotides coding for the portion of an antibody capable of binding to the target operably linked to a promoter that will permit expression of the antibody in the cell(s) of interest. The antibody is then expressed intracellularly and binds to the target, thereby disrupting the target from its normal actions.
Claims101
A method for the intracellular binding of a target antigen which comprises:
- (a) intracellular delivery of a nucleotide sequence containing a promoter operably linked to an antibody gene capable of binding to the target antigen;
- (b) intracellular expression of the antibody, wherein said antibody is intracellularly expressed as a functional antibody where said function is determined by the ability to bind to the target antigen, and wherein said antibody is selected from the group of antibodies consisting of single chain antibodies, single domain heavy chain and Fab;
- and
- (c) intracellular binding of the target antigen by said antibody.
A method for the intracellular binding of a target antigen, comprising:
- (a) introducing an antibody cassette into a cell, wherein said antibody cassette contains a nucleic acid segment encoding a light chain of an antibody and a nucleic acid segment encoding a heavy chain of an antibody operably linked to at least one promoter wherein the antibody cassette encodes a single chain antibody or Fab';
- (b) intracellular expression of said antibody encoded by said nucleic acid segments encoding said light chain and said heavy chain;
- and
- (c) intracellular binding of said target antigen by said antibody.
A method for the intracellular binding of a target antigen, which comprises:
- (a) delivery of a nucleic acid segment encoding a single chain antibody and a promoter operably linked to said nucleic acid segment to the interior of a cell;
- (b) intracellular expression of said single chain antibody;
- and
- (c) intracellular binding of said target antigen by said single chain antibody.
A method for the intracellular binding of a target antigen, which comprises:
- (a) delivery of a nucleic acid segment containing a promoter operably linked to an antibody gene capable of binding to said target antigen, wherein the antibody expressed by said antibody gene does not have a secretory sequence, and wherein the antibody is a single chain antibody or a Fab';
- (b) intracellular expression of said antibody in a form capable of binding to said target antigen;
- and
- (c) intracellular binding of said target antigen by said antibody.
A method for binding a target protein by an antibody inside a human cell at a specified location which comprises:
- (a) delivery to said cell of a nucleic acid segment encoding at least a variable light chain and a variable heavy chain of an antibody which will bind to said protein, wherein said nucleic acid segment also encodes a localization sequence,
- (b) intracellular expression of said antibody in a form capable of binding to said target protein, wherein said antibody is a single chain or Fab',
- (c) intracellular delivery of said antibody to a site directed by said localization sequence, and
- (d) intracellular binding of said target protein at said site.
A method for the intracellular binding of a target antigen, comprising:
- (a) introducing a nucleic acid segment encoding at least the heavy chain variable sequence of an antibody operably linked to a promoter into an animal cell, wherein the antibody is selected from the group consisting of single domain heavy chain, single chain, and Fab';
- (b) intracellular expression of said heavy chain variable sequence;
- and
- (c) intracellular binding of said target antigen by said heavy chain variable sequence.
A method for the intracellular binding of a target antigen, comprising:
- (a) delivery of a nucleic acid segment containing a promoter operably linked to an antibody gene encoding an antibody capable of binding to said target antigen wherein the antibody contains a secretory signal and further contains an intracellular retention sequence;
- (b) intracellular expression of an antibody encoded by said nucleic acid segment;
- and
- (c) intracellular binding of said target antigen by said antibody.
Dependencies are inferred from the claim text.