IP Library Granted Patent US 10,428,093
Granted Patent B2
US 10,428,093 · App. 15/137,448 · Granted Oct 1, 2019

Lipid co-factor essential for cell density signaling

Inventor: Richard I. Schwarz (Oakland, CA)
C07F9/572B01D61/145B01D61/16C07K14/465C07K14/47C07K14/4705C07K16/18
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Quick Facts
Patent No.
US 10,428,093
App. No.
15/137,448
Granted
Oct 1, 2019
Kind
B2
Abstract

Herein is described a cell density signaling protein and its tissue-specific lipid cofactor. Constructs, compositions and methods for isolation, purification and use of the protein and the tissue-specific lipid cofactor are described.

Claims (10)

1. An isolated tissue-specific lipid co-factor having a formula C63H123N2O11P and a mass of about 1114.89 MW, said lipid cofactor binds to a polypeptide factor that comprises the amino acid sequence set forth in SEQ ID NO: 3 or SEQ ID NO: 5 to form a protein/lipid cell density signaling complex, said cell density signally complex controlling cell proliferation, differentiation or apoptosis in bone and cartilage tissue, and said lipid co-factor comprising a ceramide phosphate, a glycerol lipid with a single fatty acid, and a linker.

2. An isolated tissue-specific lipid co-factor having a formula C30H74NO5P and a mass of about 696.65 MW, said lipid cofactor binds to a polypeptide factor that comprises the amino acid sequence set forth in SEQ ID NO: 3 or SEQ ID NO: 5 to form a protein/lipid cell density signaling complex; said cell density signally complex controlling cell proliferation, differentiation or apoptosis in tendon tissue and said lipid co-factor comprising two fatty acid chains connected directly to a phosphate molecule.

3. A method of isolating the lipid co-factor of claim 1 or 2 comprising:

a. filtering conditioned medium containing said protein lipid complex through a 0.2 micron filter;

b. adding 5M NaCl in an amount equivalent to 1/10 th the resulting conditioned medium volume;

c. using an ultrafilter in a stirred cell under pressurized nitrogen to concentrate the high salt conditioned medium to 1 ml of retentate;

d. diluting the retentate to 10 ml with water and reconcentrating to 0.5 ml;

e. capturing the free protein factor in the retentate; and,

f. capturing the free lipid cofactor in the filtrate.

4. A cell density signaling complex comprising a tissue-specific lipid cofactor of claim 1 or 2 , which is tightly bound to a polypeptide factor that comprises the amino acid sequence set forth in SEQ ID NO: 3 or SEQ ID NO: 5.

Assignments (1)
CONFIRMATORY LICENSE Recorded Jul 7, 2016
From: CALIFORNIA UNIVERSITY OF
To: ENERGY, UNITED STATES DEPARTMENT OF
Reel/Frame 039303/0224 →
Continuity (2)
Provisional Application 61896604 · Oct 28, 2013
Related Publication 20160251386A1 · Sep 1, 2016