Merck + Co., Inc.

Other

Ronald W Ellis

Paul M Keller

Robert S Lowe

Andrew J Davison

Peter J Kniskern

Arpi Hagopian

Loren D Schultz

Mark W Riemen

Donna Lorraine Montgomery

Henry Z Markus

Edward M Scolnick

Kathryn J Hofmann

Elliott D Kieff

Michael F Tuite

Hagopian Arpi

Robert B Freedman

James E Hopper

Disclosed is a process for increasing the yield of disulfide bonded recombinant proteins produced by yeast, especially recombinant secreted proteins The enzyme protein disulfide isomerase (PDI) catalyzes the formation of disulfide bonds in secretory and cell-surface proteins. We disclose the construction of recombinant strains of the yeast,which overproduce either human PDI or yeast PDI in a regulated fashion. These strains show greatly increased secretion of disulfide bonded proteins of potential therapeutic significance. These strains have the potential to increase the production of various disulfide bonded proteins.

Method of increasing production of disulfide bonded recombinant proteins by saccharomyces cerevisiae

A vaccine strain of varicella-zoster virus (VZV), tested in clinical trials, is capable of preventing chickenpox in children. This virus has been modified by the introduction into its genome of heterologous DNA which encodes an immunogenic polypeptide of another human pathogen. This heterologous polypeptide is expressed in cells infected by the recombinant virus. Such recombinant VZV is useful as a vaccine for chickenpox as well as for heterologous pathogens.

Varicella-zoster virus as a live recombinant vaccine

A gene of varicella-zoster virus (VZV) which encodes immunogenic outer surface viral proteins has been identified by DNA sequence analysis. Antibodies directed against peptides imputed from the DNA sequence can react with the glycoprotein, which itself is reactive with neutralizing antibodies. The amino-terminal sequence of the purified glycoprotein is identical to a portion of the amino acid sequence imputed from the DNA sequence. This glycoprotein is useful for the preparation of a vaccine against VZV.

DNA encoding immunogenic gpIII glycoprotein of varicella-zoster virus

The entire hepatitis B virus pre-S antigen gene linked in one contiguous reading frame to the hepatitus B virus surface antigen gene has been expressed in Saccharomyces cerevisiae. The expressed protein aggregates into a particulate form which displays the major antigenic sites encoded by both domains, thereby highlighting the utility of yeast as a host for the expression of pre-S domains. This protein is useful in in vitro diagnostic systems and as a vaccine for the treatment and prevention of hepatitis B virus-induced diseases and/or infections.

Vaccines comprising yeast-derived hepatits B virus polypeptides

The GAL4 protein is rate-limiting in quantity as a positive regulator for galactose-inducible promoters in strains of yeast. Novel strains are described in which the GAL4 protein can be overproduced in a regulatable fashion. These strains are useful for the regulatable expression in yeast of heterologous genes whose expression is driven by a galactose-inducible promoter.

Trains of yeast for the expression of heterologous genes

The hepatitis B virus preS2 antigen gene linked in one contiguous reading frame to the hepatitis B virus surface antigen gene has been expressed in Saccharomyces cerevisiae utilizing an optimized plasmid construction. The expressed protein aggregates into a particulate form which displays the major antigenic sites encoded by both domains, thereby highlighting the utility of yeast as a host for the high level expression of the preS2 as well as the S domain. This protein is useful in in vitro diagnostic systems and as a vaccine for the treatment and prevention of hepatitis B virus-induced diseases and/or infections.

Method for producing hepatitis B virus proteins in yeast

Vaccine against varicella-zoster virus

The entire hepatitis B Virus pre-S antigen gene linked in one contiguous reading frame to the hepatitis B virus surface antigen gene has been expressed in Saccharomyces cerevisiae. The expressed protein aggregates into a particulate form which displays the major antigenic sites encoded by both domains, thereby highlighting the utility of yeast as a host for the expression of pre-S domains. This protein is useful in in vitro diagnostic systems and as a vaccine for the treatment and prevention of hepatitis B virus-induced diseases and/or infections.

A gene of varicella-zoster virus (VZV) which encodes immunogenic outer surface viral proteins has been identified by DNA sequence analysis. This gene can hybrid select messenger RNA which encodes and expresses a protein which reacts with human convalescent zoster sera and with polyclonal monospecific antisera which neutralize viral infectivity. These proteins are useful for the preparation of a vaccine for VZV.

A gene of varicella-zoster virus (VZV) which encodes immunogenic outer surface viral proteins has been identified through DNA sequence analysis. Fragments of the DNA have been cloned into a vector which, when placed into a host organism, expresses proteins which react with human convalescent zoster sera and with monoclonal antibodies which neutralize viral infectivity. These proteins are useful for preparation of a vaccine for VZV.

A gene of varicella-zoster virus (VZV) which encodes immunogenic outer surface viral proteins has been identified by DNA sequence analysis. This gene can hybrid select messenger RNA which encodes and expresses a protein which reacts with human canvalescent zoster sera and with polyclonal monospecific antisera which neutralize viral infectivity. These proteins are useful for the preparation of a vaccine for VZV.

Growing community of inventors

Overbrook Hills, PA, United States of America

Ronald W Ellis