Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Retinal pigmented epithelium derived neurotrophic factor

a pigmented epithelium and neurotrophic factor technology, applied in the field of retina pigmented epithelium derived neurotrophic factor, can solve problems such as retinal degeneration

Inactive Publication Date: 2007-12-06
NAT INST OF HEALTH REPRESENTED BY THE SEC OF THE DEPT OF HEALTH & HUMAN SERVICES NAT INST OF HEALTH
View PDF12 Cites 4 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been consistently observed that loss of contact of the neural retina with the RPE of many vertebrates (retinal detachment) results in degeneration of the retina.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Examples

Experimental program
Comparison scheme
Effect test

example 1

Establishment of RPE Cell Cultures

[0151] RPE cells were harvested from post-mortem human eyes, as described by Pfeffer et al., J. Cell. Physiol. 117 333-341 (1983). The cells were grown in 75 cm flasks with Eagles's minimal essential medium supplemented with 15% v / v fetal calf serum and 5% v / v CO2 at 37° C. Cells were grown to confluence, harvested by trypsinizing the cell monolayer, and resuspended in MEM supplemented with 15% v / v fetal calf serum. A portion of the cells (about 5%) were reseeded into new 75 cm, where the cells were again grown in 5% v / v CO2 at 37° C.

example 2

Preparation of RPE-Conditioned Medium

[0152] Confluent cultures of RPE cells were washed extensively with HBSS, before conditioning, to remove serum proteins. Twenty-five ml of serum-free MEM was added, and the cultures were incubated with 5% v / v CO2 at 37° C. for about 48 hours. The conditioned medium was then collected and centrifuged at 1,000 rpm at room temperature, to remove any free cells and other particulate matter, to provide an impure PEDF protein fraction.

example 3

Purification of PEDF by SDS Gel Electrophoresis

[0153] Proteins contained in human fetal RPE-conditioned medium, prepared as described in Example 2, and in control non-conditioned medium were analyzed on an SDS-polyacrylamide slab gel.

[0154] The conditioned and non-conditioned media samples were mixed with an electrophoresis sample buffer (62.5 mM Tris, pH 6.8, 2% w / v SDS, 10% v / v glycerol, 0.001% w / v bromophenol blue, and 0.1 M 2-mercaptoethanol). Molecular-weight markers, such as phosphorylase B, bovine serum albumin, ovalbumin, carbonic anhydrase, soybean trypsin inhibitor and lysozyme (such as that supplied by Bio-Rad), were also mixed with an electrophoresis sample buffer. All the samples were denatured at 100° C., in a boiling water bath, for 5 minutes and loaded onto a SDS containing 7.5% w / v polyacrylamide gel. The electrophoresis was conducted at 20 mA until the bromphenol blue marker dye migrated to the bottom of the gel.

[0155] At the completion of electrophoresis, strip...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

PropertyMeasurementUnit
Fractionaaaaaaaaaa
Fractionaaaaaaaaaa
Percent by massaaaaaaaaaa
Login to View More

Abstract

The present invention relates to a purified retinal pigmented epithelium derived neurotrophic factor composition and a method for purifying such a retinal pigmented epithelium neurotrophic factor. The present invention also relates to a recombinant DNA molecule comprising a gene encoding a retinal pigmented epithelium derived neurotrophic factor having the DNA sequence or the amino acid sequence in SEQ ID NO:1 and to an organism transformed with the recombinant DNA molecule. In addition, the present invention relates to a method of treating tumors, ocular diseases, nerve injuries, and conditions resulting from the activity of serine proteases, which comprises administering PEDF.

Description

RELATED APPLICATIONS [0001] This application is a continuation in part of U.S. patent application Ser. No. 07 / 894,215, filed Jun. 4, 1992 and U.S. patent application Ser. No. 07 / 952,796, filed Sep. 24, 1992, which are incorporated herein by reference.[0002] This invention was made with government support under grant EY04741, awarded by The National Institutes of Health. The United States government has certain rights in this invention.FIELD OF THE INVENTION [0003] This invention relates to the purification and use of a retinal pigmented epithelium derived neurotrophic factor (PEDF). This invention also relates to a truncated version of PEDF that is referred to as PEDF-BH. In addition to PEDF and PEDF-BH and functionally equivalent proteins, this invention relates to nucleic acids that encode PEDF, PEDF-BH and functionally equivalent proteins, to vectors comprising such nucleic acids, to host cells into which such vectors have been introduced, and to the use of these host cells to pr...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
IPC IPC(8): A61K38/18C07K14/475A61K38/00C07K14/81
CPCA61K38/00C07K14/811C07K14/475
Inventor TOMBRAN-TINK, JOYCESTEELE, FINTAN R.CHADER, GERALD J.BECERRA, SOFIA PATRICIAJOHNSON, LINCOLN V.RODRIGUEZ, IGNACIO R.
Owner NAT INST OF HEALTH REPRESENTED BY THE SEC OF THE DEPT OF HEALTH & HUMAN SERVICES NAT INST OF HEALTH
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com