Cetus Corporation

Hoffmann-la Roche Inc.

Leland Stanford Junior University

David F Mark

Leo S Lin

Alice M Wang

Danute E Nitecki

Lois Aldwin

Margaret Moreland

Abla A Creasey

Corey H Levenson

Irwin Braude

Shi-da Yu Lu

Henry Rapoport

Michael V Doyle

Martha B Ladner

Janelle N Van Arsdell

James W Larrick

Kirston E Koths

Robert F Halenbeck

Ernest S Kawasaki

Shi-Da Y Lu

Gordon M Ringold

Michael W Konrad

Michael P McGrogan

Henry Rapaport

Danutee Nitecti

Frank M Torti

The present invention provides a method for determining the amount of a target acid segment in a sample by polymerase chain reaction. The method involves the simultaneous amplification or the target nucleic acid segment and an internal standard nucleic acid segment. The amount of amplified DNA from each segment is determined and compared to standard curves to determine the amount of the target nucleic acid segment present in the sample prior to amplification. The method is especially preferred for determining the quantity of a specific mRNA species in a biological sample. Additionally, an internal standard is provided useful for quantitation of multiple mRNA species.

Quantitation of nucleic acids using the polymerase chain reaction

Succinylacetone derived or related medicaments and methods of synthesis of the same are shown wherein the medicaments consists of succinylacetonyl-proline-PEG, succinylacetonyl-NH-PEG, or compounds that have the formula: ##STR1## wherein n=1-6

Immunosuppressive analogues and derivatives of succinylacetone

The present invention provides a method for determining the amount of a target acid segment in a sample by polymerase chain reaction. The method involves the simultaneous amplification of the target nucleic acid segment and an internal standard nucleic acid segment. The amount of amplified DNA from each segment is determined and compared to standard curves to determine the amount of the target nucleic acid segment present in the sample prior to amplification. The method is especially preferred for determining the quantity of a specific mRNA species in a biological sample. Additionally, an internal standard is provided useful for quantitation of multiple mRNA species.

Succinylacetone derived or related medicaments and methods of synthesis of the same are shown wherein the medicaments consists of succinylacetonyl-proline-PEG, succinylacetonyl-NH-PEG, or compounds that have the formula: ##STR1## and that have immunosuppressive activity both in vivo and in vitro based on their activities in cellular immunologic assays and adjuvant induced arthritis in rats, respectively.

A biologically active reference therapeutic protein is protected against oxidation by a method involving substituting a conservative amino acid for each methionyl residue susceptible to chloramine T or peroxide oxidation, wherein additional, non-susceptible methionyl residues are not so substituted. The oxidation-resistant mutein so produced is preferably a human mutein of interleukin-2 or interferon-.beta., and the conservative amino acid is most preferably alanine.

Oxidation-resistant muteins of Il-2 and other protein

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.

Cysteine-depleted muteins of biologically active proteins

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.

Human IL-2 like polypeptides, DNA sequences and recombinant DNA

Immunosupressive analogues and derivatives of succinylacetone

Structural genes, plasmids and transformed cells for producing cysteine

New multiclass hybrid interferon polypeptides, their corresponding encoding recombinant DNA molecules and transformed hosts which produce the new interferons are described. The amino acid sequences of these hybrids include at least two different subsequences, one of which has substantial homology with a portion of a first class of interferon (e.g., HuIFN-.alpha.) and the other which has substantial homology with a portion of a second class of interferon (e.g., HuIFN-.beta.). Data indicates the interferon activity of .alpha.-.beta. hybrids may be substantially restricted to either cell growth regulatory activity or antiviral activity.

Multiclass hybrid interferons

A modified IFN-.beta. is provided wherein the cysteine residue at position 17 is deleted and serine is substituted therefor. DNA sequences coding for the modified protein, nucleotide primers used for the mutagenesis, appropriate cloning vectors, host organisms transformed with the vectors, methods for the production and use of the modified IFN-.beta. (IFN-.beta..sub.ser17) are also provided. The specific activity of IFN-.beta..sub.ser17 is found to be substantially the same as that of native IFN-.beta..

A method for controlling weight by suppressing the normal metabolism of adipose tissue is disclosed. Administration of tumor necrosis factor (TNF) or a pharmaceutical composition containing it results in suppression of anabolism of adipose cells.

Use of tumor necrosis factor as a weight regulator

Human tumor necrosis factor (TNF) has been prepared using recombinant methods. A human promyelocytic leukemia cell line has been induced using an improved induction procedure, and the TNF purified to homogeneity. Methods, vectors, and cells useful in obtaining human TNF in practical amounts are disclosed. Muteins having N-terminal deletions, which muteins have superior biological activity, are also disclosed.

Human tumor necrosis factor

Human Tumor necrosis factor (TNF) has been prepared using recombinant methods. A human promyelocytic leukemia cell line has been induced using an improved induction procedure, and the TNF purified to homogeneity. Methods, vectors, and cells useful in obtaining human TNF in practical amounts are disclosed. Muteins having N-terminal deletions, which muteins have superior biological activity, are also disclosed.

Growing community of inventors

Danville, CA, United States of America

David F Mark