Glial cell line-derived neurotrophic factor
First Claim
Patent Images
1. A method for obtaining substantially purified glial cell line-derived neurotrophic factor protein, comprising:
- culturing cells which are transformed to express a neurotrophic factor protein selected from the group consisting of;
(a)
space="preserve" listing-type="tabular">Ser Pro Asp Lys Gln Met Ala Val Leu Pro Arg Arg Glu - Arg Asn Arg Gln Ala Ala Ala Ala Asn Pro Glu Asn Ser - Arg Gly Lys Gly Arg Arg Gly Gln Arg Gly Lys Asn Arg - Gly Cys Val Leu Thr Ala Ile His Leu Asn Val Thr Asp - Leu Gly Leu Gly Tyr Glu Thr Lys Glu Glu Leu Ile Phe - Arg Tyr Cys Ser Gly Ser Cys Asp Ala Ala Glu Thr Thr - Tyr Asp Lys Ile Leu Lys Asn Leu Ser Arg Asn Arg Arg - Leu Val Ser Asp Lys Val Gly Gln Ala Cys Cys Arg Pro - Ile Ala Phe Asp Asp Asp Leu Ser Phe Leu Asp Asp Asn - Leu Val Tyr His Ile Leu Arg Lys His Ser Ala Lys Arg - Cys Gly Cys Ile of SEQ ID NO;
6; and
(b) a neurotrophic factor protein having at least the following properties of SEQ ID NO;
6(i) a specific activity of at least about 12,000 TU/ug;
(ii) a molecular weight on reducing SDS-PAGE of about 20-23 kD;
(iii) a molecular weight on non-reducing SDS-PAGE of about 31-42 kD; and
(iv) a specific activity of at least about 24,000 times greater than the specific activity of B49-conditioned medium; and
isolating the neurotrophic factor protein.
1 Assignment
0 Petitions
Accused Products
Abstract
A novel neurotrophic factor referred to as glial cell line-derived neurotrophic factor (GDNF) has been identified and isolated from serum free growth conditioned medium of B49 glioblastoma cells. Rat and human genes encoding GDNF have been cloned and sequenced. A gene encoding GDNF has been subcloned into a vector, and the vector has been used to transform a host cell in order to produce biologically active GDNF in a recombinant DNA process.
-
Citations
11 Claims
-
1. A method for obtaining substantially purified glial cell line-derived neurotrophic factor protein, comprising:
-
culturing cells which are transformed to express a neurotrophic factor protein selected from the group consisting of; (a)
space="preserve" listing-type="tabular">Ser Pro Asp Lys Gln Met Ala Val Leu Pro Arg Arg Glu - Arg Asn Arg Gln Ala Ala Ala Ala Asn Pro Glu Asn Ser - Arg Gly Lys Gly Arg Arg Gly Gln Arg Gly Lys Asn Arg - Gly Cys Val Leu Thr Ala Ile His Leu Asn Val Thr Asp - Leu Gly Leu Gly Tyr Glu Thr Lys Glu Glu Leu Ile Phe - Arg Tyr Cys Ser Gly Ser Cys Asp Ala Ala Glu Thr Thr - Tyr Asp Lys Ile Leu Lys Asn Leu Ser Arg Asn Arg Arg - Leu Val Ser Asp Lys Val Gly Gln Ala Cys Cys Arg Pro - Ile Ala Phe Asp Asp Asp Leu Ser Phe Leu Asp Asp Asn - Leu Val Tyr His Ile Leu Arg Lys His Ser Ala Lys Arg - Cys Gly Cys Ile of SEQ ID NO;
6; and
(b) a neurotrophic factor protein having at least the following properties of SEQ ID NO;
6(i) a specific activity of at least about 12,000 TU/ug; (ii) a molecular weight on reducing SDS-PAGE of about 20-23 kD; (iii) a molecular weight on non-reducing SDS-PAGE of about 31-42 kD; and (iv) a specific activity of at least about 24,000 times greater than the specific activity of B49-conditioned medium; and
isolating the neurotrophic factor protein. - View Dependent Claims (2, 3, 4)
-
-
5. A method for obtaining a substantially purified glial cell line-derived neurotrophic factor protein, comprising the steps of:
-
(a) culturing cells which are transformed to express a neurotrophic factor protein, comprising an amino acid sequence of SEQ ID NO;
6; and(b) isolating the neurotrophic factor protein. - View Dependent Claims (6, 7, 8, 9, 10, 11)
-
Specification