Methods For The Delivery Of Toxins Or Enzymatically Active Portions Thereof
AssigneeTrustees of Tufts College
InventorsGeorge A. Oyler, Charles B. Shoemaker, Chuehling Kuo
Life of this patent
Estimating the term…
Abstract
The present invention relates to methods, systems, and kits for intoxicating cells, neuronal and non-neuronal cells, with a toxin or fragment thereof. This is done by subjecting toxin substrate and a lipid or polymeric carrier (e.g., DNA uptake facilitating agent) to one or more cells for use in cell based assays. In an aspect, the methods of the present invention allow for high throughput assays and, as such, for the evaluation of drug candidates.
Claims27
A method of intoxicating a cell with a toxin, or toxin enzymatically active fragment thereof, wherein the method comprises:
- a. mixing the toxin or toxin enzymatically active fragment thereof with a lipid or polymeric carrier;
- and
- b. exposing the mixture to one or more cells;
- wherein the cell becomes intoxicated with said toxin or toxin enzymatic fragment.
A method of delivering a botulinum neurotoxin or an enzymatically active fragment thereof to the inside of one or more cells in vitro, the method comprises:
- a. contacting the botulinum neurotoxin or an enzymatically active fragment of the toxin with a lipid or polymeric carrier to thereby obtain a mixture;
- and
- b. exposing the cells to the mixture in an amount to allow the botulinum neurotoxin or enzymatically active fragment thereof to enter the cell;
- wherein the cells are intoxicated with the botulinum neurotoxin or enzymatically active fragment thereof.
A method of preparing one or more cells for a cell-based assay, wherein the cell is intoxicated with a toxin, an enzymatically active fragment thereof;
- the method comprises:
- a. contacting the botulinum neurotoxin or an enzymatically active fragment of the toxin with a lipid or polymeric carrier to thereby obtain a mixture;
- and
- b. exposing cells to the mixture in an amount to allow the botulinum neurotoxin or enzymatically active fragment thereof to enter the cell;
- wherein the cell is intoxicated with the toxin or fragment.
A method of assessing an effect of a molecule or compound on a cell in vitro intoxicated with a toxin, an enzymatically active fragment thereof;
- the method comprises:
- a. contacting the botulinum neurotoxin or an enzymatically active fragment of the toxin with a lipid or polymeric carrier to thereby obtain a mixture;
- b. exposing cells to the mixture in an amount to allow the botulinum neurotoxin or enzymatically active fragment thereof to enter the cell;
- c. subjecting the intoxicated cells to the molecule or compound to be assessed;
- and
- d. assessing the effect on the intoxicated cells of said molecule or compound.
A system or kit for delivering a toxin, an enzymatically active fragment thereof, or a recombinant SNARE endoprotease, to the inside of a cell in vitro, wherein the system or kit comprises:
- a. one or more toxins, enzymatically active fragments thereof, or recombinant SNARE endoproteases;
- and
- b. one or more lipid or polymeric carriers.
An isolated cell that comprises:
- a toxin, an enzymatically active fragment thereof, or a recombinant SNARE endoprotease, wherein the cell comprises HEK293, HIT-T15, a neuroendocrine derive cell line, an immortalized cell line, or a tumor derived cell line.
Dependencies are inferred from the claim text.