Neovascularization inhibitors and methods for their production and use
AssigneeThe Johns Hopkins University
InventorBert M. Glaser
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Abstract
Neovascularization inhibitors are disclosed, which are purified polypeptides recovered from cultured cells, including retinal pigment epithelial cells and human fibroblast cells. The polypeptides may be used for the treatment of diseases in which new blood vessel formation plays a role, such as diabetic retinopathy, senile macular degeneration, tumor growth, and rheumatoid arthritis.
Claims13
A process for producing a neovascularization inhibitor comprising the steps of:
- culturing retinal pigment epithelial cells to produce a culture medium containing said neovascularization inhibitor;
- and
- purifying said neovascularization inhibitor from said medium wherein said neovascularization inhibitor has a molecular weight of approximately 57,000.+-.3,000 and an isoelectric point of approximately 4.6.+-.0.3 and is stable in an environment with a pH of approximately from 2 to 3.
A process for producing a neovascularization inhibitor comprising the steps of:
- culturing retinal pigment epithelial cells to produce a culture medium containing said neovascularization inhibitor;
- separating said medium from said cells;
- and
- substantially purifying said inhibitor by subjecting said medium to hydrophobic interaction chromatography wherein said neovascularization inhibitor has a molecular weight of approximately 57,000.+-.3,000 and an isoelectric point of approximately 4.6.+-.0.3 and is stable in an environment with a pH of approximately from 2 to 3.
A process for obtaining a neovascularization inhibitor produced by retinal pigment epithelial cells comprising the steps of:
- culturing retinal pigment epithelial cells to produce a culture medium containing a neovascularization inhibitor wherein said neovascularization inhibitor has a molecular weight of approximately 57,000.+-.3,000 and an isoelectric point of approximately 4.6.+-.0.3 and is stable in an environment with a pH of approximately from 2 to 3;
- separating said medium from said cells;
- bringing said medium into contact with immobilized antibodies to said neovascularization inhibitor so as to bind said neovascularization inhibitor in said medium to said immobilized antibodies;
- and
- separating said neovascularization inhibitor from said immobilized antibodies.
A process for producing a neovascularization inhibitor comprising the steps of:
- growing human retinal pigment epithelial cells in a culture medium;
- separating said medium from said cells;
- subjecting said separated medium to acid dialysis to produce a dialysate containing said inhibitor;
- substantially purifying said inhibitor in said dialysate by reverse-phase high performance liquid chromatography;
- and
- purifying said substantially purified inhibitor by gel permeation high performance liquid chromatography wherein said neovascularization inhibitor has a molecular weight of approximately 57,000.+-.3,000 and an isoelectric point of approximately 4.6.+-.0.3 ad is stable in an environment with a pH of approximately from 2 to 3.
Dependencies are inferred from the claim text.