This section will provide information on worldwide patents relevant to vaccine design and production. The Patent Report will give the following information: title of patent, patentee, patent number, publication and summary of the patent. A number of patents in this report are reproduced from 'Biotechnology Abstracts' with permission of Derwent Publications Ltd

Hepatitis B virus vaccine production from hepatitis B surface antigen: purification by polyethylene glycol precipitation isopyknic flotation and gel chromatography

Thomssen W. German 3150-935; 30 June 1983

In a new process for the production of hepatitis B virus vaccine from hepatitis B surface antigen (HBsAg) from the blood of HBsAg carriers, the 20 nm particles are prepurified by precipitation with 6% polyethylene glycol, enriched with 12% polyethylene glycol, and purified by flotation in a salt solution of density 1.34-1.20 g/ml, followed by gel chromato- graphy on agarose. In a new procedure for inactivation of residual infectivity of the hepatitis B vaccine, the inactivating agent (0.4 mg/ml formaldehyde) is removed by washing the HBs antigen adsorbed on aluminium hydroxide gel. The simple 3-stage purification gives 5000-fold enrichment, and yields a product of 99% purity. Higher formalin concen- trations can be used in the inactivation step, and removal of the inactivating agent by washing immobilized antigen avoids the need for dialysis which is difficult to perform under sterile conditions. 001-84

Recombinant DNA synthesizing surface antigen of B type hepatitis virus: insertion in the metabolic enzyme gene of vector DNA

Jpn Gan unexamined; 30 May 1983

Recombinant DNA synthesizing a hepatitis B virus surface antigen, in which a DNA fragment containing a surface antigen gene of hepatitis B virus DNA as a DNA-donor, e.g. DNA of the serum, tissue or cells of patients with hepatitis B virus, (e.g~ subtype ado is inserted in a metabolic enzyme gene of vector DNA. The hepatitis B virus DNA is cut by BamHI to separate a fragment containing a surface antigen gene. The DNA fragment is inserted into vector DNA. The recombinant DNA is cut by TaqI to separate the surface antigen gene. The DNA fragment is inserted into a metabolic enzyme gene of pBR322. The recombinant DNA, integrated in Escherichia coil produces the surface antigen in large amounts. The DNA fragment containing the surface antigen is separated and reacted with T4 DNA-polymerase. pBR322 DNA cut by PstI is reacted with SI nuclease. The two DNA fragments are ligated with T4 DNA-ligase to give hybrid DNA. Ca-treated E. coli is infected with the hybrid DNA and posistive colonies are detected by a colony hybridization method 002-84

Vucciniu virus modified with exogenous DNA in vaccinia genome: useful for vaccination of animals to form antibodies to coded antigens

Health Res Eur. 83-286; 6 July 1983

Vaccinia virus modified by the presence of exogenous DNA in the vaccinia genome is new. Biologically pure cultures of

Potent Report vaccinia virus VTK-79 (ATCC VR 2031) and VTK-79L are new. Immunization of a host animal susceptible to vaccinia virus is effected by inoculation with a vaccinia virus having, within the vaccinia genome, DNA exogenous to the genome, usually from plasmid pBR322, and coding for an antigen that induces the animal to develop antibodies against it. Human and animal individuals and populations may have missing genes or genetic material introduced for modification. replacement or repair or defective genes. In vivo recombina- tion ofvaccinia virus and donor DNA involves contact of a cell monolayer with a cell-compatible medium containing the vaccinia virus and donor DNA which comprises DNA for introduction into vaccinia genome present in a DNA segment. 003-84

Anti-plogue vaccine prepared from Yersiniapestis EV strain by extraction of cellular membrane antigen and purification and fractionation

L'Air Liquide Fr. 2517-966; 17 June 1983

The process for the production of an anti-plague (bubonic plague) biochemical vaccine from Yersinia pestis strain EV comprises 5 steps. (a) Fermentation of Y pest/s in liquid medium at a pH slightly above neutral, under medium aerobiosis, at 29 to 31°C for a period depending on the inoculum. (b) Low speed centrifugation, lysis of the bacterial culot, the higher speed centrifugation of the lysate and separation of the culot of cellular membranes. (c) Extraction of the vaccinating membrane antigens by contacting the cellular membrane culot with an aqueous solution of sodium chloride under continuous shaking at refrigerated tempera- ture (0 to 4°C) and pH 8.0 for70 h. (d) Low speed centrifugation and decantation of the membrane extract, followed by purification of the supernatant extract by ultrafdtration using a filter membrane to give the desired MW cut. (e) Dialysis of the lowest fractions and lyophilization of vaccinating antigenic fractions. This vaccine is free from secondary reactions and more stable and consistent than living vaccines. 01)4-84

Lyophilized duck hepatitis ,accine from sspltn sta'als virus: by cultivation in chicken embryos followed by homogenization preservation and lyophilization

Phylaxia UK 2111-829; 13 July 1983

Lyophilized duck hepatitis vaccine is produced by inoculating the alantoic cavity of SPF-chicken embryos with attentuated hepatitis virus TIN, Asplin strain and then incubating at 37°C. Embryos which die between 24 and 96 h after inoculation are recovered, beaks and eyes removed and the remainder homogenized in physiological saline. The eggs must be specifically pathogen free. The homogenizing mixture can contain from 65 to 75% by vol. physiological saline plus stabilizers. Before iyophilization the vaccine is checked for sterility. The supernatant from centrifugation is treated with preservatives and stabilizers e.& antibiotics. Then it is lyophilizcd at -30 to -40°C. This vaccine can be stored for up to a year at 4°C, whereas previously vaccine could only be stored for 6 months at -20°C. 005-84

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