ANALOGUE HIN47 OF HAEMOPHILUS WITH REDUCED PROTEASE ACTIVITY Russian patent published in 2003 - IPC

FIELD: immunology, microbiology, genetic and protein engineering. SUBSTANCE: invention relates to immunogens and antigens of Haemophilus species. Isolated and purified analogue of Hin47 protein from Haemophilus influenzae exhibits decreased protease activity that is less 10% of activity of natural protein Hin47 but the same immunogenic properties as natural protein due to substitution or incorporation of at least one amino acid in native protein determining protease activity. Amino acid can be taken among amino acids 195-201, His-91 or Asp- 121 of native protein Hin47. Chimeric molecule comprises analogue of protein Hin47 of Haemophilus influenzae bound with polysaccharide. Recombinant plasmid DS-1011-1-1 (pT7/Hin47*) carries nucleic acid molecule expressing analogue of protein Hin47 with substitution of Ser-197 for alanine. At least a single amino acid residue of protein Hin47 determining protease activity is identified. For removal or substitution of nucleotide sequence encoding this amino acid site-directed mutagenesis of gene hin47 is carried out. Mutant gene is incorporated into cell to obtain transformed cell and the latter is cultured to yield analogue of Hin47. Immunogenic composition comprises effective amount of analogue Hin47. Interaction of sample with analogue Hin47 is carried out to obtain complexes comprising analogue Hin47 and recognizing its antibodies specifically if they present in sample. In subject immunization antibodies exhibiting specificity to protein Hin47 are obtained. Interaction of sample with antibodies is carried out. Diagnostic set for assay of antibodies presence that are able to recognize protein Hin47 specifically comprises in sample analogue of protein Hin47 and agents providing interaction of analogue with sample. Invention ensures to obtain vaccine preparations against sicknesses caused by pathogenic microorganism Haemophilus influenzae. EFFECT: improved method of preparing, valuable biological and medicinal properties. 18 cl, 25 dwg, 3 tbl, 14 ex