Merck + Co., Inc.

Other

Henry Z Markus

William J McAleer

Kathrin Ute Jansen

Loren D Schultz

Michael P Neeper

Ronald W Ellis

Joseph G Joyce

Kathryn J Hofmann

Donna Lorraine Montgomery

Hugh A George

James C Cook, Iii

Roy A Machlowitz

Fred J Bailey

Robert Z Maigetter

Mark Rosolowsky

Robert B Freedman

Michael F Tuite

Ernest Dale Lehman

Loren S Schultz

Synthetic DNA molecules encoding the HPV31 L1 protein are provided. Specifically, the present invention provides polynucleotides encoding HPV31 L1 protein, wherein said polynucleotides are free from internal transcription termination signals that are recognized by yeast. Also provided are synthetic polynucleotides encoding HPV31 L1 wherein the polynucleotides have been codon-optimized for high level expression in a yeast cell. The synthetic molecules may be used to produce HPV31 virus-like particles (VLPs), and to produce vaccines and pharmaceutical compositions comprising the HPV31 VLPs. The vaccines of the present invention provide effective immunoprophylaxis against papillomavirus infection through neutralizing antibody and cell-mediated immunity.

Optimized expression of HPV 31 L1 in yeast

Synthetic DNA molecules encoding the HPV31 L1 protein are provided. Specifically, the present invention provides polynucleotides encoding HPV31 L1 protein, wherein said polynucleotides are free from internal transcription termination signals that are recognized by yeast. Also provided are synthetic polynucleotides encoding HPV31 L1 wherein the polynucleotides have been codon-optimized for high level expression in a yeast cell. The synthetic molecules may be used to produce HPV31 virus-like particles (VLPs), and to produce vaccines and pharmaceutical compositions comprising the HPV31 VLPs. The vaccines of the present invention provide effective immunoprophylaxis against papillomavirus infection through neutralizing antibody and cell-mediated immunity 

Optimized expression of HPV31 L1 in yeast

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

Recombinant expression vectors encoding the L1 and L2 proteins of papillomavirus, methods of making and using the recombinant proteins and purified virus-like particles comprised of the recombinant proteins are provided.

Recombinant papillomavirus vaccines

The invention comprises a lyophilized live herpes virus vaccine that comprises from about 0.5% to about 8% moisture.

Stable lyophilized live herpes virus vaccine

Hepatitis B surface antigen (HBsAg) is produced in vitro in high titer and purity from tissue cultures of cells that shed HBsAg. The cells are grown on hollow fiber capillary units having a molecular weight cut-off of about 10,000.

Methods of producing HBsAg

Hepatitis A virus is harvested from persistently infected cells, the cells are then trypsinized, replanted and a second harvest of hepatitis A virus is obtained. The cells can be trypsinized and replanted and harvested indefinitely.

Process for preparing hepatitis A virus

An improved stabilized liquid live viral vaccine contains a live virus, partially hydrolyzed gelatin, a monosaccharide or disaccharide, a cell culture medium, L-glutamic acid, L-arginine and sufficient physiologically acceptable acidic buffer to maintain the pH at from about 6.0 to about 6.5.

Vaccine stabilizer containing L-glutamic acid and L-arginine

Hepatitis A virus (HAV) is produced in high titer from tissue cultures grown on a hollow fiber capillary unit.

High titer production of hepatitis A virus

Hepatitis B surface antigen (HB.sub.s Ag) is produced in vitro in high titer and purity from tissue cultures of cells that shed HB.sub.s Ag by growing the cells with a first incubation at elevated temperature followed by a second incubation at a lower elevated temperature.

Method of producing hepatitis B surface antigen

Automated tests to evaluate the antibody titer of a sample by incubating a test sample, bacteria, and complement, and detecting bacterial growth in the incubated mixture, and to identify bacteria and its titer in a sample by incubating a test sample, bacterial antiserum, and complement, and detecting inhibition of bacterial growth in the incubated mixture. The automated tests lend themselves to diagnostic use and may be provided in the form of diagnostic test kits.

Bacterial or bacterial antibody assay

Lyophilization time of live viral compositions is reduced and output per lyophilization unit is increased by lyophilizing a reduced volume of a more concentrated viral composition.

Lyophilization process for live viral compositions

A stabilized lyophlized or liquid live viral vaccine contains a live virus, partially hydrolyzed gelatin, a 6-carbon polyhydric alcohol, Medium 199 and acidic buffer.

Growing community of inventors

Wyncote, PA, United States of America

Henry Z Markus