Production of bacterial extracts for the manufacture of vaccines by ultrasonic lysis of ultrafiltered bacterial cells

Patent Report from the whole blood. The vaccine and hyperimmune serum are useful for immunizing mammals and birds against diseases caused by enterotoxin-producing Gram-negative bacteria. The immune modulator described is a detoxified extract of lipopolysaccharide and is useful in combination with many antigens to enhance the primary immune response. 039-86

separated from the medium and expanded. The expanded cells are then grown in medium containing hypoxanthine and azaserine. The monoclonal antibodies produced are effective for the treatment of tetanus in mammals. They can be used passively to immunize individuals who suffer from tetanus or who are at risk with respect to tetanus infection. (/42-86

Non-A non-B hepatitis surface antigen useful for the preparation of vaccines and methods of use - purification of non-A non-B surface antigens from serum or plasma

Escherichia coli GaI-Gal pilus protein or fragments

Inst. Merieux US 4542 016; 17 September 1985 A new vaccine against non-A non-B hepatitis virus comprising non-A non-B surface antigen (NANBs antigen) is described which contains NANBs antigen in a physiologically acceptable medium. The vaccine is prepared by selecting serum or plasma in which the presence of NANBs antigen has been demonstrated, and purifying the latter. The purification process may be performed by e.g. affinity chromatography, ultracentrifugation, gel chromatography, precipitation with e.g. PEG, or ultrafiltration. To prepare the vaccine, the purified antigen is dissolved in a physiologically acceptable medium comprising an apyrogenic buffer, e.g. phosphate buffer (pH 7). An adjuvant may also be added, and the vaccine may be a composite vaccine. Anti-NANBs antibodies which could serve to detect the presence of NANBs antigens in man can be prepared. 040-86 Production of bacterial extracts for the manufacture of vaccines by ultrasonic lysis of ultrafiltered bacterial cells

Bruschettini Belgian 902 735; 16 October 1985 The production of total bacterial extracts for vaccine production is claimed. The bacteria are rendered virulent in vivo and are cultured in vitro and concentrated by ultrafiltration in a closed system using a membrane with a mol.wt, cut off of 100 000. The cells are then subjected to ultrasonic lysis using a continuous flow chamber with Ti probes supplied at 1l)0() W and emitting at 22 Hz. The bacteria are rendered virulent by injecting the bacteria into the pleural cavity of a rabbit, and the pleural exudate is recovered after 20 h. This method gives high purity extracts with high antigenic activity. In an example, Staphylococcus aureus in heart-brain broth was diluted with Mellin broth and injected into the pleural cavity of a rabbit. After 20 h, the pleural cavity was aspirated, plated out and selected for virulence. The virulent colonies were inoculated with Difco BH broth and incubated at 37°C for 18 h and then for a further 24 h with the addition of more broth. The bacteria were concentrated on a Millipore HPCF filter and the retentate was sonicated. The lyzate was diluted with phenol to give a vaccine. (/41-86 Human lymphoblastoid cell line for hybridoma generation and monoclonal antibody production for tetanus treatment

C~,tus Eur. 160 550; 6 November 1985 A human lymphoblastoid B cell of the line LTR228 and its progeny, including ouabain-resistant progeny, is claimed. A method, for the production of human lymphoblastoid B cells such as LTR228, which have the ability to fuse with human B lymphocyte to produce human X human hybridomas, comprises culturing Epstein-Barr virus transformed human B lymphoblastoid cells, preferably derived from the WI-L2 line in a medium containing 6-thioguanine, and selecting from the resulting 6-thioguanine cells those which efficiently fuse with normal human lymphocytes. Monoclonal antibody producing human hybridomas are prepared by fusing lymphoblastoid B cells of the line LTR228 with human antibody producing cells in a fusion medium containing a fusogen. The cells are 132

Vaccine, Vol. 4, June 1986

Vaccines against urinary tract infections containing new

Leland-Stanford-Yr. Univ. EP 161 095; 13 November 1985 Pilus vaccines for treating urinary tract infections in humans are claimed. These contain a polypeptide with an amino acid sequence corresponding to at least one antigenic determinant of Gal-Gal pilus protein. The Escherichia coli HU849 GalGal pilus protein comprises a sequence of 163 amino acids, and the 79-110, 15-7(I, 133 63 and 11 t-25 fragments of this protein are new. This protein may be obtained by isolation and purification from Escherichia coli HU849 pili, peptide synthesis or recombinant DNA technology using the appropriate DNA coding sequence. In an example, a suspension of E. coli HU849 cells was sheared at 4000 rev. min i for 30 rain at 4°C to separate the pill which were repeatedly reprecipitared from Tris/NaCl by adding MgC12. Dialysis against water gave the pilus protein with a purity of 97-99% and a lipopolysaccharide content of less than 0.1%. These fragments are highly effective and specific in generating antibodies to urinary pathogens and are obtainable in practical amounts and in pure form. (/43-86 Continuous cultivation of bacteria for vaccine production with continuous harvesting using a centrifuge; application to e.g.

Bordetella bronchiseptica Inst. Immunpraep. Naehrmed. E. German 225 707; 7 August 1985 Continuous cultivation of bacteria for the production of whole-cell vaccines in human and veterinary medicine is effected using a fermenter which is operated continuously under aseptic conditions and is connected to a continuousflow centrifuge. This also operates under aseptic conditions. With this apparatus, high productivities can be achieved. In an example, an 80 litre bubble column fermentor was charged with 70 litres of a casein hydrolysate medium and inoculated with Bordetella bronchiseptica. Aeration was affected at 4~ 5 1 min i for 12-15 h, and the fermenter was then supplied with 10-15 1 h i culture medium while transferring an equal amount of broth via a peristaltic pump through a sterile hose to a continuous-flow centrifuge. Over 24 h, 2603(10 litres of broth with an average cell count of 1.5 × 1011/ml were processed, corresponding to a dry cell yield of 1000-1200 g. 044-86 Antigenic protein material with adenylate-cyclase activity isolated from Bordetella pertussis and used in vaccines against whooping cough with reduced activity

Wellcome Eur. 162 639; 27 November 1985 A vaccine composition for conferring protection against Bordetella pertussis includes an antigenic preparation derived from B. pertussis and comprising proteinaceous material associated with adenylate-cyclase (EC 4.6.1.1) activity (ACAP), together with a carrier. The isolation of an antigenic preparation containing ACAP comprises treating a culture of B. pertussis cells with an aq. amino acid buffer at pH 2.5-3.5. The cells are separated and the preparation is isolated from the remaining fluid. Vaccines containing ACAP confer protection against whooping cough. In an example, the crude outer membrane protein was isolated from B. pertussis cells isolated 3/4 of the way through the exponential