Compositions and methods for induction and modulation of angiogenesis and methods and assays for identifying angiogenesis modulators

Patent metadata

Field	Value
Publication number	US-9821013-B2
Application number	US-201414243123-A
Country	US
Kind code	B2
Filing date	Apr 2, 2014
Priority date	Apr 2, 2013
Publication date	Nov 21, 2017
Grant date	Nov 21, 2017

Abstract

The present disclosure provides methods and compositions, including a placental extract, for inducing and/or modulating angiogenesis; methods of identifying modulators of angiogenesis, and assays for identifying modulators of angiogenesis. The present disclosure also provides methods of making a composition, including a placental extract that can induce and/or modulate angiogenesis.

First claim

The invention claimed is:

1. A method of making a human placental extract, the method comprising:
obtaining a sample from a human placenta;
removing blood from the placental sample to produce a crude placental extract;
mixing the crude placental extract with a protein solubilization agent to solubilize and linearize proteins in the crude extract;
separating and removing solid materials from the solubilized protein-placental extract;
performing dialysis on the solubilized protein-placental extract to remove the protein solubilization agent; and
after dialysis, removing remaining solids from the extract to produce the human placental extract, whereby the resulting human placental extract contains less than 2% normalized protein abundance of collagen IV or laminin.

2. The method of claim 1 , wherein removing blood from the placental sample comprises homogenizing the human placenta sample with a buffer, centrifuging the homogenized sample, and discarding the supernatant containing blood.

3. The method of claim 1 , wherein the protein solubilization agent is urea.

4. The method of claim 1 , wherein separating solid materials from the solubilized protein-placental extract comprises centrifuging the solubilized protein-placental extract and removing solids.

5. The method of claim 1 , wherein the method is conducted at a temperature between about −86° C. and about 5° C.

6. The method of claim 1 , wherein removing remaining solids from the dialyzed protein-placental extract comprises centrifuging the dialyzed protein-placental extract and removing remaining solids.

7. The method of claim 1 , wherein the resulting human placental extract contains less than 2% normalized protein abundance of laminin.

8. A method of making a human placental extract, the method comprising:
obtaining a sample from a human placenta;
removing blood from the placental sample to produce a crude placental extract;
mixing the crude placental extract with a protein solubilization agent to solubilize and linearize proteins in the crude extract;
separating and removing solid materials from the solubilized protein-placental extract;
performing dialysis on the solubilized protein-placental extract to remove the protein solubilization agent; and

after dialysis, removing remaining solids from the extract to produce the human placental extract, whereby the resulting human placental extract contains less than 2% normalized protein abundance of collagen IV.

9. A method of making a human placental extract, the method comprising:
obtaining a sample from a human placenta;
removing blood from the placental sample to produce a crude placental extract;
mixing the crude placental extract with a protein solubilization agent to solubilize and linearize proteins in the crude extract;
separating and removing solid materials from the solubilized protein-placental extract;
performing dialysis on the solubilized protein-placental extract to remove the protein solubilization agent; and

after dialysis, removing remaining solids from the extract to produce the human placental extract, whereby the resulting human placental extract contains less than 2% normalized protein abundance of collagen IV and less than 2% normalized protein abundance of laminin.

Assignees

• Univ Florida

Inventors

• Mcfetridge Peter S
• Moore Marc C

Classifications

• A61P43/00

  Drugs for specific purposes, not provided for in groups A61P1/00-A61P41/00 · CPC title

• A61K38/18

  Growth factors; Growth regulators · CPC title

• A61L2300/252

  Polypeptides, proteins, e.g. glycoproteins, lipoproteins, cytokines · CPC title

• C12N2502/1347

  Smooth muscle cells · CPC title

• A61L27/3808

  Endothelial cells · CPC title