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5196321 Methods for in vitro cleavage of ubiquitin fusion proteins
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PATENT ABSTRACTS
5196320 METHOD OF PRODUCING ENGINEERED BINDING PROTEINS Stephen D Gillies assigned to Abbott Biotech Inc Disclosed are methods of producing fusion proteins including those with dual biological activities. These methods include the provision of a first and second D N A sequence encoding a first and second polypeptide, repectively, the digestion of the first D N A sequence at a restriction site adjacent its 3' or 5' terminus, and the ligation of a linker/adapter sequence (l/a) to the restricted end of the first D N A sequence, thereby forming a cassette. The l/a includes, at one end, that portion of the first DNA sequence extending from its terminus nearest the restriction site to the restriction site, and at the other end, one side of a splice site. A eucaryotic host cell is transfected with the cassette and the second D N A sequence having, at one end, one side of a splice site compatible with the side of the splice site on the l/a. The transfected host cell is cultured to express the transfected DNA as a single chain fusion protein.
5196322 VECTORS AND COMPOUNDS FOR EXPRESSION OF ZYMOGEN FORMS OF HUMAN PROTEIN C Nils U Bang, Hartmut Ehrlich, Brian W Grinnell, S Betty Yan, Cologne, assigned to Eli Lilly and Company A method for the recombinant production of zymogen forms of human protein C is described. These zymogen forms differ from native zymogen protein C in their increased sensitivity to activation by thrombin and thrombin/thrombomodulin. DNA compounds, vectors, and transformants useful in the method are also disclosed.
5196323 PROCESS FOR PREPARING AND PURIFYING ALPHA-INTERFERON 5196321 METHODS FOR IN VITRO CLEAVAGE OF UBIQU1TIN FUSION PROTEINS Andrea Bachmair, Daniel Finley, Alexander Varshavsky, Cologne, Federal Republic Of Germany assigned to Massachusetts Institute of Technology Methods of designing or modifying protein structure at the protein or genetic level to produce specified amino-termini in vivo or in vitro are described. The methods can be used to alter the metabolic stability and other properties of the protein or, alternatively, to artificially generate authentic amino-termini in proteins produced through artificial means. The methods are based upon the introduction of the use of artificial ubiquitin-protein fusions, and the discovery that the in vivo half-hfe of a protein is a function of the amino-terminal amino acid of the protein.
Gerhard Bodo, Ingrid Maurer-Fogy, Edgar Falkner, Silvia J Lindner, Cologne, assigned to Boehringer Ingelheim International GmbH A process for the preparation and purification of recombinant Interferon- alpha is disclosed. The invention is directed to a process comprising the following steps: cultivating E. coli containing the interferon gene for a growth period during which not more than 5 ~ methionine-interferon is formed; extracting and concentrating the expressed interferon; subjecting the preliminarily purified material to Tandem Chromatography, wherein the Tandem Chromatography comprises separation on a cellulose column followed by an antialpha-interferon monoclonal antibody affinity column; subjecting the thus purified material to isoelectric precipitation of impurities at about pH 4.0 to about pH 4.8; and purifying the interferon by chromatography on a high performance cation exchange column using a volatile buffer, wherein the purified interferon is nonimmunogenic when administered parenterally to a human.
PATENT ABSTRACTS
5196320 METHOD OF PRODUCING ENGINEERED BINDING PROTEINS Stephen D Gillies assigned to Abbott Biotech Inc Disclosed are methods of producing fusion proteins including those with dual biological activities. These methods include the provision of a first and second D N A sequence encoding a first and second polypeptide, repectively, the digestion of the first D N A sequence at a restriction site adjacent its 3' or 5' terminus, and the ligation of a linker/adapter sequence (l/a) to the restricted end of the first D N A sequence, thereby forming a cassette. The l/a includes, at one end, that portion of the first DNA sequence extending from its terminus nearest the restriction site to the restriction site, and at the other end, one side of a splice site. A eucaryotic host cell is transfected with the cassette and the second D N A sequence having, at one end, one side of a splice site compatible with the side of the splice site on the l/a. The transfected host cell is cultured to express the transfected DNA as a single chain fusion protein.
5196322 VECTORS AND COMPOUNDS FOR EXPRESSION OF ZYMOGEN FORMS OF HUMAN PROTEIN C Nils U Bang, Hartmut Ehrlich, Brian W Grinnell, S Betty Yan, Cologne, assigned to Eli Lilly and Company A method for the recombinant production of zymogen forms of human protein C is described. These zymogen forms differ from native zymogen protein C in their increased sensitivity to activation by thrombin and thrombin/thrombomodulin. DNA compounds, vectors, and transformants useful in the method are also disclosed.
5196323 PROCESS FOR PREPARING AND PURIFYING ALPHA-INTERFERON 5196321 METHODS FOR IN VITRO CLEAVAGE OF UBIQU1TIN FUSION PROTEINS Andrea Bachmair, Daniel Finley, Alexander Varshavsky, Cologne, Federal Republic Of Germany assigned to Massachusetts Institute of Technology Methods of designing or modifying protein structure at the protein or genetic level to produce specified amino-termini in vivo or in vitro are described. The methods can be used to alter the metabolic stability and other properties of the protein or, alternatively, to artificially generate authentic amino-termini in proteins produced through artificial means. The methods are based upon the introduction of the use of artificial ubiquitin-protein fusions, and the discovery that the in vivo half-hfe of a protein is a function of the amino-terminal amino acid of the protein.
Gerhard Bodo, Ingrid Maurer-Fogy, Edgar Falkner, Silvia J Lindner, Cologne, assigned to Boehringer Ingelheim International GmbH A process for the preparation and purification of recombinant Interferon- alpha is disclosed. The invention is directed to a process comprising the following steps: cultivating E. coli containing the interferon gene for a growth period during which not more than 5 ~ methionine-interferon is formed; extracting and concentrating the expressed interferon; subjecting the preliminarily purified material to Tandem Chromatography, wherein the Tandem Chromatography comprises separation on a cellulose column followed by an antialpha-interferon monoclonal antibody affinity column; subjecting the thus purified material to isoelectric precipitation of impurities at about pH 4.0 to about pH 4.8; and purifying the interferon by chromatography on a high performance cation exchange column using a volatile buffer, wherein the purified interferon is nonimmunogenic when administered parenterally to a human.