Abstract: Stable pharmaceutical compositions suitable for parenteral administration to mammals are prepared which are in a pH range of about 2 to about 4 and comprise a therapeutically effective amount of a recombinant interferon-.beta. protein (IFN-.beta.) dissolved in an inert carrier medium comprising as a stabilizer/solubilizer an effective amount either of glycerol or of polyethylene glycol polymers having an average molecular weight from about 190 to about 1600 daltons. Further disclosed and claimed are methods for extracting IFN-.beta. from a microbial host transformed to produce it and then purifying and formulating said IFN-.beta. and methods for screening for other polyhydric non-detergent stabilizer/solubilizers or combinations thereof as solubilizer/stabilizers for pharmaceutical compositions of IFN-.beta..
Abstract: A stable, continuous human cell line or progeny thereof is produced that is resistant to 6-thioguanine and ouabain, secretes less than 40 ng/ml of endogenous IgM antibodies, and grows with a doubling time of about 18 hours. The cell line, which preferably is adapted to serum-free medium, may be used as a fusion partner with an antibody-producing cell line so as to generate antibodies. In addition, it may be electroporated with a vector containing a gene of interest to produce a transformed cell line which generates a protein encoded by the gene, such as an IgG or IgM antibody.
Abstract: The invention discloses modified signal peptides derived from wild-type signal peptides of the type that are capable of forming membrane-bound lipoproteins and methods for making such modified signal peptides and DNA sequences encoding them. Modified signal peptides of the invention and DNA sequences encoding them are useful for increasing the secretion of heterologous gene products produced by transformed host organisms. The invention further discloses a method for producing recombinant DNA sequences in vivo.
Abstract: An improved process for recovering and purifying lipophilic recombinant proteins such as human .beta.-interferon and interleukin-2 from their hosts yields a protein preparation which may be formulated into a stable pharmaceutical composition having a therapeutically effective amount of the biologically active recombinant lipophilic protein dissolved in a non-toxic, inert, therapeutically compatible aqueous-based carrier medium at a pH of 6.8 to 7.8 which medium also contains a stabilizer for the protein, such as human serum albumin, normal serum albumin and human plasma protein fraction.
Type:
Grant
Filed:
September 13, 1985
Date of Patent:
February 12, 1991
Assignee:
Cetus Corporation
Inventors:
Wolfgang H. Hanisch, Peter M. Fernandes, Terrance Taforo
Abstract: Damage to cells, tissue and other body parts in a mammalian host may be treated by using a lymphokine or cytotoxin in conjunction with at least one biological modifier, which may be a free radical scavenger or a metabolic inhibitor. The lymphokine or cytotoxin is preferably tumor necrosis factor and the biological modifier is preferably uric acid, buthionine sulphoximine, vitamin C, aspirin, or nordihydroguaiaretic acid. Such a combination may be used to treat, for example, cancer, infectious diseases, and damage caused by radiation therapy, high oxygen tension, and chemotherapy.
Type:
Grant
Filed:
August 25, 1989
Date of Patent:
January 15, 1991
Assignee:
Cetus Corporation
Inventors:
Robert Zimmerman, Benedict J. Marafino, Jr.
Abstract: DNA constructs are prepared which operably link human interferon genes, selective, eukaryotic marker genes, and promoter and expression control sequences for the expression of human interferon in Chinese hamster ovary (CHO) cells or progeny thereof. The human recombinant interferon so produced contains glycans which are a subset of the population of glycans which are contained in the native counterpart, and may be used in therapeutic formulations. The CHO cells yield high levels of human interferon with no detectable amounts of host IFN, either constitutive or inductive.
Type:
Grant
Filed:
July 31, 1985
Date of Patent:
October 30, 1990
Assignee:
Cetus Corporation
Inventors:
Francis P. McCormick, Michael A. Innis, Gordon M. Ringold
Abstract: DNA plasmids are described which are selected mutants in which an altered repressor gene leads to high copy number replication. Elements in the plasmids are modified in such a way that readthrough expression of heterologous DNA inserted in the plasmid will not continue into the replication primer strand. Deletions resulting from interference with replication primer strand transcription are thereby avoided.
Abstract: Oligonucleotides derivatized to incorporate horseradish peroxidase (HRP) are useful in diagnostic methods and provide significant advantage over radioactively labeled oligonucleotide probes. The HRP-derivatized oligonucleotides can be easily and efficiently prepared by reacting a sulfhydryl containing oligonucleotide with a conjugate of a mal-sac-HNSA ester and HRP under mild and easily controlled reaction conditions.
Type:
Grant
Filed:
November 16, 1989
Date of Patent:
October 9, 1990
Assignee:
Cetus Corporation
Inventors:
Corey Levenson, Chu-an Chang, Fred T. Oakes
Abstract: The invention herein is directed to methods using Procion dyes to perform separations of interest in manipulating the NAD.sup.+ -independent ribotoxins. The methods are useful for preparing therapeutic agents containing these ribotoxins or their A polypeptide components. This separation method has been applied in particular to preparing hybrid toxins containing ricin toxins, both for purifying the resulting products and also for separating the components intended to be used in the preparation of these end products. In addition, a novel ricin isotoxin prepared using the method of the invention is disclosed.
Abstract: Conjugates comprising recombinant ricin toxin A chain and a binding moiety which may be an antigen binding portion selected from the group consisting of Fab, Fab' and F(ab').sub.2 fragments of monoclonal or polyclonal antibodies, hormones, lectins or other compounds that are recognized by cell receptors, are described and claimed.
Abstract: An approved procedure for the purification and renaturation of biologically active, bacterially produced IFN-.beta. is described. The partially purified material obtained by solubilization of isolated refractile bodies from the recombinant cells is treated to obtain reduction of the protein in the presence of a chaotropic environment and then oxidized after removal of the reducing agent. However, the chaotropic environment is retained during the oxidation. Upon removal of the chaotropic environment, a solubilizing additive is supplied to maintain the IFN-.beta. in solution. Further purification by conventional means may also be effected.
Type:
Grant
Filed:
May 11, 1987
Date of Patent:
October 9, 1990
Assignee:
Cetus Corporation
Inventors:
Susan Hershenson, Ze'ev Shaked, Jody Thomson
Abstract: Muteins of biologically active proteins such as IFN-.beta. and IL-2 in which cysteine residues that are not essential to biological activity have been deleted or replaced with other amino acids to eliminate sites for intermolecular crosslinking or incorrect intramolecular disulfide bridge formation. These muteins are made via bacterial expression of mutant genes that encode the muteins that have been synthesized from the genes for the parent proteins by oligonucleotide-directed mutagenesis.
Type:
Grant
Filed:
February 7, 1985
Date of Patent:
September 25, 1990
Assignee:
Cetus Corporation
Inventors:
David F. Mark, Leo S. Lin, Shi-da Yu Lu, Alice M. Wang
Abstract: Immunotoxins comprising a cytotoxic moiety and monoclonal antibodies which bind to human ovarian cancer tissue having at least one of the following capabilities are claimed: cytotoxic ID.sub.50 of 10 nM or less against human ovarian cancer cells, retardation of human ovarian cancer tumor growth in mammals or extension of survival of a mammal carrying a human ovarian cancer tumor. Antigens to which the monoclonal antibody of the immunotoxin bind are identified and characterize the immunotoxins. Methods of killing human ovarian cancer cells, retarding the growth of human ovarian cancer tumors in mammals or extending the survival of mammals carrying human ovarian cancer tumors are claimed.
Type:
Grant
Filed:
July 6, 1987
Date of Patent:
September 18, 1990
Assignee:
Cetus Corporation
Inventors:
Michael J. Bjorn, David J. FitzGerald, Arthur E. Frankel, Walter J. Laird, Ira H. Pastan, David B. Ring, Mark C. Willingham, Jeffrey L. Windelhake
Abstract: Immunotoxins comprising a cytotoxic moiety and an antigen binding portion selected from the group consisting of Fab, Fab' and F(ab').sub.2 fragments of a monoclonal antibody, which binds to human ovarian cancer tissue, having one of the following capabilities are claimed: cytotoxic ID.sub.50 of about 10 nM or less against human ovarian cancer cells, retardation of human ovarian cancer tumor growth in mammals, or extension of survival of a mammal carrying a human ovarian cancer tumor. Antigens or epitopes to which the monoclonal antibodies bind are identified and characterize the immunotoxins. In a preferred embodiment an immunotoxin comprising at least an antigen binding portion of a monoclonal antibody, which binds to human transferrin receptor, but does not block binding of transferrin to the receptor, is described and claimed. Immunotoxin comprising the F(ab').sub.2 region of the antitransferrin monoclonal antibody are also claimed.
Type:
Grant
Filed:
July 6, 1987
Date of Patent:
September 11, 1990
Assignee:
Cetus Corporation
Inventors:
Michael J. Bjorn, Arthur E. Frankel, Walter J. Laird, David B. Ring, Jeffrey L. Winkelhake
Abstract: Coupling of biological materials is disclosed via 4-hydroxy-3-nitrobenzene sulfonic acid sodium salt (HNSA) esters. The novel activated esters are structured so as to react at one end with an amine group of a selected biological or non-biological material, thereby releasing the spectroscopically monitorable HNSA dianion, and at the other end with, typically, a sulfhydryl group of a second material which may or may not be biological. The esters provide for a cross-linking reaction that may be easily controlled and monitored.
Abstract: A method and a cloning vector are described for the controlled accumulation of cloned heterologous gene products in Bacillus subtilis. The cloning vector is capable of being replicated in B. subtilis and includes the heterologous gene located and oriented such as to be under the control of an operator, promoter, and ribosome binding site sequence. The gene codes for a protein which is under the control of a transport mechanism by which the protein is secreted by the B. subtilis. The gene product is recovered from the growth medium for the B. subtilis. The cloning vector is also capable of similar use in other bacteria such as E. coli.
Type:
Grant
Filed:
June 19, 1987
Date of Patent:
August 28, 1990
Assignee:
Cetus Corporation
Inventors:
Shing Chang, Hing C. Wong, Vaughan P. Wittman
Abstract: Recombinant diphtheria toxin A fragment muteins which are enzymatically inactive but immunologically crossreactive with diphtheria toxin are disclosed. Intermediates and methods for preparing such proteins using recombinant techniques are also described.
Type:
Grant
Filed:
March 7, 1988
Date of Patent:
August 21, 1990
Assignee:
Cetus Corporation
Inventors:
Lawrence Greenfield, Anne W. Emerick, Walter J. Laird
Abstract: A system for expression of coding sequences for desired heterologous proteins in procaryotic hosts whereby the protein is produced intracellularly in soluble, biologically active form, is disclosed. The expression is obtained by ligation of the coding sequence downstream of, and proximally to, but out of reading frame with, the terminated leader encoding sequence for a secreted bacterial protein, such as alkaline phosphatase. The resultant proteins are influenced by the leader sequence codons to effect the desired three-dimensional conformation, but not to effect secretion.
Type:
Grant
Filed:
March 17, 1988
Date of Patent:
August 14, 1990
Assignee:
Cetus Corporation
Inventors:
Michael Piatak, Jr., Walter J. Laird, Julie A. Lane
Abstract: Coupling of biological materials is disclosed via 4-hydroxy-3-nitrobenzene sulfonic acid sodium salt (HNSA) esters. The novel activated esters are structured so as to react at one end with an amine group of a selected biological or non-biological material, thereby releasing the spectroscopically monitorable HNSA dianion, and at the other end with, typically, a sulfhydryl group of a second material which may or may not be biological. The esters provide for a cross-linking reaction that may be easily controlled and monitored.
Abstract: DNA sequences and recombinant DNA molecules which encode human IL-2 IL-2 like polypeptides, hosts transformed with vectors carrying the DNA sequences as inserts, methods for their production and therapeutic formulations utilizing the IL-2 and IL-2 like polypeptides are provided.
Type:
Grant
Filed:
August 23, 1988
Date of Patent:
July 3, 1990
Assignee:
Cetus Corporation
Inventors:
Michael P. McGrogan, Ernest S. Kawasaki, Michael V. Doyle, David F. Mark