Merck + Co., Inc.

William J Miller

William J McAleer

William M Hurni

Peter J Kniskern

Arpi Hagopian

Edward H Wasmuth

Dennis J Kubek

Pamela D Burke

John P Hennessey

Charlotte C Ip

Richard L Tolman

Stephen Marburg

Maurice R Hilleman

Philip J Provost

Raymond E Spier

Alexander U Bertland, Ii

Roy A Machlowitz

Lynn T Callahan

John P Hennessey Jr

Type-specific capsular polysaccharide preparations from Streptococcus pneumoniae, having on average less than about 1000 oligosaccharide repeat units per molecule, polydispersities between 1.0 and 1.4, intrinsic viscosities between 0.6 and 3.0 dL/g, and less than 3% contamination of type-specific polysaccharide by group-specific C-polysaccharide, are produced by a novel process. The novel type specific polysaccharide products are useful in the preparation of vaccines, especially covalent conjugates comprising the novel polysaccharide linked to a T-cell stimulatory carrier protein. Vaccines comprising the novel polysaccharides are useful in the prevention of infection and of diseases associated with infection by Streptococcus pneumoniae.

Process for making capsular polysaccharides from Streptococcus

A novel conjugate vaccine comprising partially hydrolyzed, highly purified, capsular polysaccharide (Ps) from Streptococcus pneumoniae bacteria (pneumococci, Pn) linked to an immunogenic carrier protein, is produced by a new process. The conjugate is useful in the prevention of pneumococcal infections. Vaccines comprising a mixture of from one to ten different pneumococcal polysaccharide-immunogenic protein (Pn-Ps-PRO) conjugates induce broadly protective recipient immune responses against the cognate pathogens from which the polysaccharide components are derived. Young children and infants younger than 2 years old, normally unable to mount a protective immune response to the Pn-Ps alone, exhibit protective immune responses upon vaccination with these Pn-Ps-PRO conjugates.

Pneumococcal polysaccharide conjugate vaccine

Anti-hepatitis B virus surface protein (anti-HBS) antibody is adsorbed onto the surface of a substratum. Hepatitis B virus PreS2+S (PreS2+S) protein is then adsorbed onto the same surface through the interaction of the anti-HBS antibody with the 'S' portion of the PreS2+S protein. The coated surface is then incubated concomitantly with the test sample and a radiolabelled antibody specific for the 'PreS2' portion of the PreS2+S protein.

Radio-immuno assay for hepatitis B virus PreS2 antibodies

Hepatitis B PreS2+S antigen is isolated from plasma by adsorption on an affinity chromatography column, elution with a chaotropic agent and treatment with concentrated urea at an elevated temperature. The process retains all or substantially all of the preS2+S antigen.

Pres2+S hepatitis B vaccine derived from plasma

A quantitative assay for HAV comprises infecting at a temperature of from about 30.degree. to about 37.degree. C. a cell sheet with a dilution series of hepatitis A virus and then adding Newcastle disease virus (NDV) to the infected cell sheet and incubating the infected cell sheet in the presence of NDV at an elevated temperature for a time sufficient to permit a cytopathic effect (CPE) to become manifest, this time typically being at least about 4 days and preferably about 5 days. At an incubation temperature of from about 31.degree. to about 33.degree. C. the absence of CPE indicates the presence of HAV and the presence of CPE indicates the absence of HAV. At an incubation temperature of from about 34.degree. to about 36.degree. C. the absence of CPE indicates the absence of HAV and the presence of CPE indicates the presence of HAV.

Assay for hepatitis a virus

HBcAb in a biological fluid is adsorbed on a surface which is then coated with BSA. The coated surface is then incubated first in the sample and then in the presence of radiolabelled HBcAb.

RIA assay for HBcAg

A diagnostic reagent for hepatitis A antibody is prepared by adhering hepatitis A antibody to a surface by non-specific adsorption followed by specific coupling of hepatitis A antigen to the antibody. This reagent is useful in an in vitro assay for hepatitis A antibody.

Hepatitis A antibody assay

Semi-automated assays for viral infectivity and an assay for serum neutralizing antibody content are based on staining the cells and macroscopic reading of the cytopathic effect.

Macroscopic method for determining cytopathic effects in viral assay

An immune adherence hemagglutination assay for detection of hepatitis A antigen or antibody. Plasma having known high titer of hepatitis A antibody, and gamma globulin having known high titer of hepatitis A antibody.

Human immune serum globulin with high hepatitis A antibody titer

Liquid containing hepatitis A antibody is adsorbed on a surface. The surface is then coated with a proteinaceous material and incubated in the presence of the sample to form a complex of hepatitis A antigen and hepatitis A antibody. The complex is incubated again in the presence of excess radioactively labelled hepatitis A antibody, and the resulting radionuclide is measured.

Assay for hepatitis A antigen

HB.sub.c Ab is detected in biological fluid by contacting HB.sub.c Ag derived from Dane particles and adhered to a surface with the biological fluid and an antibody-enzyme conjugate. The HB.sub.c Ag is adhered to the surface in the presence of a nonionic surfactant.

Assay for hepatitis B core antibody

Compositions comprising HB.sub.s Ag and a physiologically acceptable medium are useful as diagnostic and immunologic agents.

Hepatitis B core antigen composition

HB.sub.c Ag is prepared by isolating Dane particles by isopycnic banding of fluid from HB.sub.s Ag positive donors, optionally but preferably pelleting the Dane particles, and then removing the surface antigen by contacting the Dane particles with a nonionic surfactant having from about 15 to about 35 oxyethylene units in the presence of a reducing agent such as mercaptoethanol. The isolated core antigen is used as a diagnostic and immunologic agent.

Method of preparing hepatitis B core antigen

Hepatitis A antigen obtained from the stools of patients infected with hepatitis A (infectious hepatitis) virus has been found to be usable in fast, simple assays for hepatitis A antibody and thus usable in diagnosis of hepatitis A disease, and for the preparation of hepatitis A vaccine.

Hepatitis A antigen

A process for the controlled manufacture of cells and vaccines in which reproducable real time quantitative assays are utilized to ascertain the physical and chemical state of the system, in order to more efficiently manage the system, so that cell sheet formation may be determined and the vaccine may be harvested at its maximum titre, thereby resulting in significantly increased yields and decreased production costs.

Growing community of inventors

North Wales, PA, United States of America

William J Miller