Method of isolation and purification of fusion polypeptides

Chemistry: molecular biology and microbiology – Micro-organism – tissue cell culture or enzyme using process... – Recombinant dna technique included in method of making a...

Patent

Rate now

  [ 0.00 ] – not rated yet Voters 0   Comments 0

Details

435 697, 435219, C12N 952, C12P 2100

Patent

active

055321429

ABSTRACT:
Methods for isolation and purification of recombinant proteins are described. Fusion proteins incorporating a cleavage site sensitive to proteolysis by a plant virus proteinase may be cleaved from carrier proteins to provide high yields of protein product. Methods employing a plant virus proteinase to cleave expressed fusion proteins are particularly suitable for obtaining proteolytically sensitive polypeptides in the presence of added cell protease inhibitors. Also disclosed are recombinant vectors useful for overproducing plant virus proteinases in a suitable host.

REFERENCES:
patent: 5162601 (1992-11-01), Slightom
patent: 5179007 (1993-01-01), Jarvis et al.
patent: 5221619 (1993-06-01), Itakura et al.
Rorrer et al., J. Gen. Virol., 73:775-783, 1992.
Allison et al., "The Nucleotide Sequence of the Coding Region of Tobacco Etch Virus Genomic RNA: Evidence for the Synthesis of a Single Polyprotein," Virology, 154:9-20, 1986.
Carrington et al., "A Second Proteinase Encoded by a Plant Potyvirus Genome," The EMBO Journal, 8(2):365-370, 1989.
Carrington and Dougherty, "A Viral Cleavage Site Cassette: Identification of Amino Acid Sequences Required for Tobacco Etch Virus Polyprotein Processing," Proceedings of the National Academy of Science, USA, 85:3391-3395, 1988.
Carrington and Dougherty, "Small Nuclear Inclusion Protein Encoded by a Plant Potyvirus Genome is a Protease," Journal of Virology, 61(8):2540-2548, 1987.
Dahlman et al., "High Level Expression in Escherichia coli of the DNA-Binding Domain of the Glucocorticoid Receptor in a Functional Form Utilizing Domain-Specific Cleavage of a Fusion Protein," The Journal of Biological Chemistry, 264(2):804-809, 1989.
Dougherty et al., "Biochemical and Mutational Analysis of a Plant Virus Polyprotein Cleavage Site," The EMBO Journal, 7(5):1281-12887, 1988.
Guan and Dixon, "Eukaryotic Proteins Expressed in Escherichia coli: An Improved Thrombin Cleavage and Purification Procedure of Fusion Proteins with Glutathione S-Transferase," Analytical Biochemistry, 192:262-267, 1991.
Montgomery et al., "Expression of an Autoprocessing CAT-HIV-1 Protinase Fusion Protein: Purification to Homogeneity of the Released 99 Residue Protinase," Biochemical and Biophysical Research Communications, 175(3):784-794, 1991.
Parks et al., "Cleavage Profiles of Tobacco Etch Virus (TEV)-Derived Substrates Mediated by Precursor and Processed Forms of the TEV NIa Proteinase," Journal of General Virology, 73:149-155, 1992.
Taylor and Drickamer, "Carbohydrate-Recognition Domains as Tools for Rapid Purification of Recombinant Eukaryotic Proteins," Biochemistry Journal, 274:575-580, 1991.
Waldenstrom et al., "Synthesis and Secretion of a Fibrinolytically Active Tissue-Type Plasminogen Activator Variant in Escherichia coli," Gene, 99:243-248, 1991.
Parks et al., Virology 182:17-27 (1991).
Uhlen, M. et al.; Meth. Enzymol. 185:129-143 (1990).
Smith, M. C. et al.; J. Biol. Chem. 263:7215 (1988).
Restrepo-Hartwig et al., J. Virol. 66:5662-5666 (1992).
Dougherty et al., Virology 172:302-310 (1989).
Dougherty et al., Virology 183:449-456 (1991).
Louis et al., Eur. J. Biochem. 199:361-369 (1991).
Menendez-Arias et al., J. Biol. Chem. 267:24134-24139 (1992).

LandOfFree

Say what you really think

Search LandOfFree.com for the USA inventors and patents. Rate them and share your experience with other people.

Rating

Method of isolation and purification of fusion polypeptides does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Method of isolation and purification of fusion polypeptides, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Method of isolation and purification of fusion polypeptides will most certainly appreciate the feedback.

Rate now

     

Profile ID: LFUS-PAI-O-1505850

  Search
All data on this website is collected from public sources. Our data reflects the most accurate information available at the time of publication.