Chemistry: natural resins or derivatives; peptides or proteins; – Proteins – i.e. – more than 100 amino acid residues – Hormones – e.g. – prolactin – thymosin – growth factors – etc.
Patent
1989-01-05
1991-04-30
Moskowitz, Margaret
Chemistry: natural resins or derivatives; peptides or proteins;
Proteins, i.e., more than 100 amino acid residues
Hormones, e.g., prolactin, thymosin, growth factors, etc.
530412, 530415, 530417, 530418, 530419, 530420, 530427, C07K 314, C07K 318, C07K 324, C07K 328
Patent
active
050119141
ABSTRACT:
A ciliary neurotrophic factor (CNTF), particularly sciatic nerve CNTF (SN-CNFT) is claimed. The SN-CNTF described herein is a single protein species and has a specific activity that increased to greater than 25,000-fold from crude extract. The purification is carried out by lowering the pH of the crude nerve extract preparation to form a precipitate which is removed and discarded; raising the pH to about 6.3 followed by ammonium sulfate fractionation; chromatofocusing a solution containing a second precipitate obtained from the 30% to 60% ammonium sulfate containing solution; subjecting the fractions obtained by chromatofocusing to sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis (PAGE); and performing reversed-phase high-performance liquid chromatography (HPLC) on the SDS-PAGE eluate.
Amino acid data for this SN-CNTF is also provided. In addition, methods for using this data for providing SN-CNTF probes and for screening cDNA and genomic libraries are also provided. Recombinant-DNA methods for the production of SN-CNTF are described.
REFERENCES:
patent: 4923696 (1990-05-01), Appel et al.
Scoper, R. K. 1987, Protein Purification; Principles and Practice, Springer Varlag, N.Y., pp. 33-35, 45-54, 93-99, 111-113, 199-208 and 285-290.
Sofer et al., 1983, Bio Techniques, Nov.-Dec., pp. 198-203.
Manthorpe et al., 1986, Brain Research 367(10:282-286.
Popiela et al., 1981, Developmental Biology 83:266-277.
Watters et al., 1987, J. of Neurochemistry 49(3): 705-713.
Mizrachi et al., 1986, i J. of Neurochemistry, 46(6): 1675-1682.
Bonneryia et al., 1986, Biol Technology 4:954-958.
Collins Franklin D.
Lin Leu-Fen
Furman Keith
Moskowitz Margaret
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