Patent Reports

which the embryos are treated with trypsin to give high quantitities of cells (more than 100 million per egg) for use in virus cultivation. Vaccines having a high virus concentration (above I million UFP/0.5 ml). The process economizes on starting material, giving the same quantity of vaccine from 1/I 0th the number of white-free eggs. The quality of the vaccines is superior to present vaccines: they are very pure and free from allergic reactions, have a virus concentration about 10 times that of known vaccines and are stable on storage. 068-84

Detoxified conjugates of haptens and toxin carrier molecules - immunogenic compositions for vaccination especially hepatitis B surface antigen coupled to diphtheria toxin

lnsL Pasteur Fr 2532-850:16 March 1984

Detoxified, conjugated molecule having immunogenic pro- perties and preferably having a vaccinating effect with respect to a determined pathogenic principle and which carries one or more haptens, which have at least one antigenic determinant char- acteristic of the pathogenic principle, is immobilized on a carrier molecule. The conjugate is particularly characterized in that the carrier molecule is a toxin derivative or a fragment of a toxin and that the haptens are immobilizied on the aminoacyl residues of the toxin other than one of the two terminal aminoacyl groups of the carrier molecule. The method produces immunogenic compositions which may be used to vaccinate against specific pathogens, especially against hepatitis B surface antigen (HBsAg). The vaccines are effective and avoid undesirable secondary reactions. Detailed examples describe the coupling of various HBsAg sequences to diphtheria toxin to prepare immunogenic compositions suitable for vaccination purposes

069-84

Vaccine for preventing diarrhoea in animals, prepared by mixing enterotoxin and velotoxin produced by Bacillus derived from pig diarrhoea with cilia and inactivation

Kitasato Re~ Inst. Jpn unexamined, 9020-226; 2 February 1984

Vaccine is obtained by mixing enterotoxin and velotoxin produced by Bacillus derived from pig diarrhoea with cilia, and .inactivating the mixture. The preparation comprises: (a) cul- turing enterotoxin and velotoxin produced Bacilli: (b)cenlri- fuging the culture and filtering the supernatant; (c) adding 0.5% formalin to the filtrate; (d) incubating at 37°C for 24 h to obtain immunogen containing enterotoxin and velotoxin; and (1) culturing cilia-containing Bacillus strain; (2) adding phosphate buffer saline; (2) after filtration, preparing a cell suspension containing 5 )< 10 I° CFU/ml; (4) adding 0.5% formalin; (4) incubating at 37°C for 24 h with shaking, to obtain the immunogen with cilia; mixing the former immunogen with the latter immunogen in equal amounts; and adding to the mixture 6-7 mg/I of aluminum gel. The inactivated vaccine is ad- ministered to mother pigs one month before delivery, the vaccine is effective in prevention of diarrhoea caused by coliform bacilli in the suckling pigs. 070-84

Streptococcus group B antigen preparation, vaccine preparation against all serotypes of group B

Re~ Corp. N Y US 4,439,422; 27 March 1984

Production of pure group B streptococcus antigen and pure type 111 streptococcus antigen of group B Streptococcu,~ spp, com- prises; (i) growth of group B type I11 Streptococcus in a nutrient medium at pH 7 and free from animal proteins and lipids. The medium contains nutrients, including I-5% glucose and 0.4-0.16 M phosphate. Growth is at 34-38°C for 12-72 h: (ii) the medium is cooled to4°C; (iii) bacterial cells are removed: (iv) the medium is dialysed against water, while buffer is added to the equivalent of 0.01 M Tris, pH 7; (v) the retentate is treated with anion exchanger (equilibrated with the buffer); (vi) the anion exchanger is eluted with a linear NaCi gradient of 0-0.75 M NaCI in the same buffer, and group B antigen is eluted at 1).02-0.15 M NaCI. The type III antigen is eluted at 0.15-0.22 M NaCI. Finally the fractions are cooled separately and applied to agarose gel

chromatography. The antigens are obtained in pure condition and are in a form suitable to prepare human vaccines for giving protection against all serotypes of group B Str~Ttococcus spp. 071-84

Vaccine against colibacillosis in young cattle and pigs, Escherichia coli ATCC 39076 obtained by conjugation; !£88 and K99 antigens

Merck-USA US 4,440,748; 3 April 1984

Biologically pure culture of Escherichia coil synthesizing both K88 and K99 antigens, deposited as ATCC 39076 is new. The strain is obtained by bacterial conjugation of a donor b,] coli (K88+, NalS, Raf+) ATCC 39075 and a recipient E. coli (K99+, NalR) A T C C 39074. The two strains are incubated together at 37°C for 8-24 h and the resulting cells are used to inoculate a nutrient medium containing raffinose as sole C-source and nalidixic acid. The cells are incubated at 37°C for 2-5 days and viable cells are separated from the medium. The vaccine is obtained by isolation of K88 and K99 antigen from this strain. This is followed by purification and use in a pilus vaccine to stimulate antibody production. The colibacillosis vaccine can either be comprised of K88 and K99 antigens or the whole inactivated cells together with a carrier. 072-84

Vaccine for stimulation of protection against hepatitis B virus - contains hepatitis B surface antigen from transformed micro- organisms

Sk + I'- RIT Aust. 8316-750:15 March 1984

Recombinant DNA molecule comprising a nucleotide sequence coding for hepatitis B surface antigen (HBsAg) and a regulatory region derived from the yeast arg3 gone to effect transcription of the HBsAg sequence in yeast is described, which can be used to produce a vaccine for stimulation of protection against hepatitis B virus (HBV) infection. The DNA fragment and the HBsAg produced by it can be used as a probe for detection of HBV in biological samples by DNA hybridization techniques and immunoassays. The HBsAG sequence is preferably positioned relative to the regulatory region such that the HBsAg synthesized by expression of the sequence is free from extraneous amino acid residues. The region is preferably derived from pYe ura3 arg3, e.g. a yeast DNA fragment specifying the arg3 gone obtained by digestion of pYe ura3 arg3 with Hindlll is used. The HBsAg sequence may be obtained by treatment of HBV DNA with BamHl. An example describes a Saccaromyce~ ccrevixiue strain obtained after transformation of the yeast with the plasmid.

073-84

Production of antigen associated with adult T cell leukaemia - by treatment of cells containing the antigen with surfactant

Eisai Eur 105-465:18 April lt)84

Production of adult T cell leukaemia-associated antigen is described and comprises treatment of cells producing the antigen with a surfactant followed by the recovery of the antigen. A method for assaying adult T cell leukaemia-associated antibodies is also described and comprises (1) provision of adult T cell leukacmia-associatcd antigen producing cells. (2) addition of sodium deoxycholate, octylphenoxypolyethoxyethanol and polyoxyethylene(10)octylphenyl ether surfactant to the cells in aqueous medium, (3) removal of the antigen and (4) immuno- assay of a human blood serum sample with the antigen. This assay is very sensitive and the highly active antigen can be recovered in high yield. The antigen producing cells are especially MT-2 cells derived from umbilical cord leukocytes from a mixture of leukaemia cells of an adult T cell leukaemic female patient and similar cells from a normal male neonate. After surfactant addition, the mixture is dialyzed and the retentate centrifuged to give an antigen containing supernatant. The immunoassay may involve enzyme labelling, radioactive labelling or passive hcmagglntination. 074-~4

288 Vaccine, Vol. 2., December 1984