phase of growth in a defined medium at 36-37°C with agi- tation, by suspending them in glycine-HCl buffer (pH 3) and sedimenting. The supernatant was separated, centrifuged, filtered and freeze-dried. The protein was purified by DEAE- Trisacryl chromatography and by preparative flat bed isoelec- tric focusing in granulated gel etc. 045-86

Production of pseudorabies virus subunit vaccines useful for conferring protection against virus infections and for diagnosis especially with sheep antisera; cloning and expression of glyco- protein genes

Mol. Genetics Eur. 162 738; 27 November 1985

Methods and compositions are described for the identifica- tion, cloning and expression of pseudorabies virus (PRV) glycoprotein genes in prokaryotic and eukaryotic expression systems. Methods for culturing these new host cell organisms to produce PRV gene products are also described. Localiza- tion of genes encoding antigen viral proteins in large DNA virus genomes is described. Vaccines containing the polypep- tide (I) having an immunoreactive and antigenic determinant of PRV protein are obtained economically and in large and useful amounts for use in conferring protection against PRV. (I) Is obtained by culturing a unicellular organism e.g. Escher- ichia coli containing a recombinant vector comprising a DNA sequence encoding it and capable of being replicated, trans- cribed and translated in the organism. The (I) is isolated from the culture. The genetically engineered PRV product may be used to produce antibodies for use in passive immunotherapy or as a diagnostic tool. 046-86

Recombinant plasmid contains DNA coding for polio virus genome for vaccine production

Nippon-Polio-Res. Inst. Japan 0207 582; 19 October 1985

A new recombinant plasmid contains DNA complementary to all the genes contained in a weak-toxicity strain of polio virus, for use in vaccine production. In an example, polio virus Sabin-1 strain was grown in HeLa cells, and polio virus RNA (35 S) was extracted, cDNA was prepared using this RNA as template, from which double-standed DNA was derived by the action of reverse transcriptase. The double- stranded DNA and plasmid pBR322 were each cleaved with restriction enzymes BamHI, HindIII and PstI. Each of the fragments derived from the double-stranded DNA was mixed with ring-opened pBR325 and linked together using DNA ligase. Eight recombinant plasmids thus obtained were spliced in a specific sequence to give the recombinant plasmid. This was introduced into HeLa cells by the calcium phosphate method, and the treated cells were incubated at 33.5°C. The polio virus obtained is identical to the mother virus used for cloning and is useful for vaccine production. 047-86

DNA coding for antigenic papilloma virus e.g. HPV IA poly- peptide for use in vaccine production and diagnosis

Inst. Pasteur US 4551 270; 5 November 1985

DNA fragments are described coding for antigenic papilloma virus, especially those of the HPV 1A type, polypeptides. Three new polypeptides (I), (II) and (III) (where X is Lys or Arg) encoded by these DNA fragments are described. These DNA sequences may be cloned by producing expression vectors and transforming suitable microorganism hosts. The peptides are useful in the production of vaccine compositions and in papilloma virus diagnosis.

L e u - A s p - G l n - P h e - P r o - L e u - G l y - A r g - X - P h e - L e u (I) A l a - L y s - A r g - A r g - A r g - L y s (II) Ala -Lys-Lys-Lys-Lys-Lys (III)

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Patent Report

Long surviving human influenza vaccine preparation by gene reassortment of a human and bird virus

St. Jude-Child. Hosp. US 4552 757; 2 November 1985

A long surviving human influenza vaccine is claimed. This is derived from a bird-human influenza reassortment virus whose non-surface protein genes are derived from a bird influenza A parent and whose surface antigen genes are derived from the human influenza parent. At least one of the bird influenza nucleoprotein and matrix protein genes are present in the reassortment. The bird-human influenza reas- sortment can be A/Washington/80 (H3N2) x A/Mallard/New York/6750/78 (H2N2), A/California/78 (HINI ) xA/Mallard/ New York/6750/78 (H2N2) or A/Udorn/307/72 x A/Mallard/ Alberta/573/78, A/Pintail/Alberta/121/79 or A/Mallard/New York/6750/78. The two viruses are mated together in a bird cell culture medium and the desired progency are selected at high temperature, using antibodies to the surface antigens of the parent bird virus. Liver viruses are obtained which can be used as vaccines which replicate efficiently in mammals, do not cause influenza in man but give long lasting protection against infection. 049-86

Hepatitis B vaccine surface antigen isolation from serum

Nat. Res. Develop. Corp. US 4554 157; 19 November 1985

A protein fraction suitable for the formulation of a vaccine against hepatitis B virus infections is produced by: (a) treating serum-derived particles of 20-25 nm diameter and having the hepatitis B surface antigen with a nonionic detergent, prefer- ably an alkaryl polyether alcohol; and (b) affinity chromato- graphy of the treatment mixture to yield a preparation con- taining an immunogenic glycopeptide of mol.wt. 28 000 (IGP) and an immunogenic polypeptide of mol.wt. 23 000 (IP). The mixture of IGP, IP and detergent is used to form a layer on, top of an aq. solution, buffered to pH 7.0-7.5 to avoid denaturation of IGP and UP, and centrifuged on a sucrose gradient of 20-50 wt.%. An immunogenic aq. frac- tion virtually free from detergent and containing micelles of IGP and IP is obtained. The buffer preferably contains Na3PO4, NaCI and EDTA. Centrifugation is performed at 40 000-250 000g at 4-20°C for about 24 h. The amount of detergent in the immunologically active fragments is suffi- ciently reduced to allow a clinically useful vaccine to be prepared. 050-86

New synthetic polypeptides useful as immunogens and in vaccines for preventing infections by leukaemia-associated viruses

Scripps World 8504 871; 7 November 1985

A synthetic polypeptide is claimed which contains about 8-40 amino acid residues in a sequence corresponding to a peptide sequence of an envelope protein antigenic determinant encoded by a leukaemia associated virus. This polypeptide has a sequence which corresponds to at least part of the envelope protein homologous to the small envelope protein p l5E of feline leukaemia virus FeLV-B (from positions 45- 70, 105-150 or 70-95 taken from the amino terminus) or the large envelope protein gp. 70 of FeLV-B (from positions 45- 65, 205-240, 290-330, 345-70 or 390-420 taken from the amino terminus). The peptide sequences of the virus envel- ope proteins are determined by recombinant DNA cloning techniques. An antigenic determinant domain is selected and the corresponding polypeptide is prepared by solid phase synthesis using a cysteine resin. These polypeptides are useful in vaccines and in immunogens for the treatment and preven- tion of infections caused by leukaemia-associated viruses.

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Vaccine, Vol. 4, June 1986 133