[The purification and concentration of the rabies virus by a diafiltration concentration method]. / Ochistka i kontsentrirovanie virusa beshenstva metodom diafil'tratsionnogo kontsentrirovaniia.

Purified concentrates of rabies virus were prepared by both microfiltration chromatography and diafiltration methods. The diafiltration concentrate had a high level of protective activity and was not inferior to the microfiltration chromatographic one. The infectious activity was better retained after diafiltration, the chromatographic purity of both preparations was similar. It is concluded that diafiltration presents a good alternative to the currently used methods for purification and concentration of viruses.

[Chemotactic function of skin fibroblasts in patients with amyloidosis]. / Khemotaksicheskaia funktsiia fibroblastov kozhi u bol'nykh amiloidozom.

Chemotaxis of cultivated fibroblasts, obtained from patients with amyloidosis, chronic glomerulonephritis and healthy volunteers, was investigated. Fibroblast migration toward donor serum and serum from patients with amyloidosis was measured using Boyden chamber's technique. As "zero" chemoattractant Hank's solution was used. It was shown, that chemotactic index (CI) was independent from cell density. Significant CI depression of fibroblasts from patients with amyloidosis toward donor serum in contrast to fibroblasts from patients with chronic glomerulonephritis and healthy volunteers was shown. The depression of chemotactic function was the same with fibroblasts from patients with different variants of amyloidosis and different stages of amyloid nephropathy and was stable in several cell generations. The results obtained suggest the existence of primary hereditary variant (variants) of chemotactic function, which may lead to the development of amyloidosis in certain conditions.

[The submolecular structure of the Carnation Mottle virus]. / O submolekuliarnom stroenii viriona Carnation Mottle Virus.

The model of the tertiary structure of the CMtV subunit was suggested and the mode of its packaging into the capsid was determined on the basis of electron microscopy small-angle X-ray scattering and X-ray diffraction data. The i-ray diffraction data on the crystal of the native virion of Carnation Mottle Virus were obtained on the automatic diffractometer supplied with two-dimensional detector.

[Use of microfiltration and gel filtration technics for purifying and concentrating the Venezuelan equine encephalomyelitis virus]. / Primenenie mikrofil'tratsionnoi i gel'-fil'tratsionnoi tekhniki dlia ochistki i kontsentrirovaniia virusa venesuél'skogo éntsefalomielita loshadei.

A system for purification and concentration of Venezuelan equine encephalomyelitis (VEE) virus omitting large-scale ultracentrifugation was developed. The first step consists in prefiltration through large pore nuclear filters (NF) to remove large particle admixtures from the virus-containing fluid. In the second step, the resulting filtrage undergoes concentrating microfiltration through small pore NF. In this way the virus, but not total protein, is concentrated approximately 30-fold without any loss of biological activity. For VEE virus purification the next step uses gel filtration chromatography in a column with chemically modified macropore silica. In this stage, the purification factor by protein is approximately 100-fold and virus yield reaches 40%. It is suggested that the procedure used be applied for purification and concentration of a wide range of enveloped viruses.

[Preparative isolation of myxovirus glycoproteins and the study of their immunogenic properties]. / Preparativnoe poluchenie glikoproteinov miksovirusov i izuchenie ikh immunogennykh svoistv.

Preparative isolation of glycoproteins from ortho- and paramyxoviruses is described. The purified concentrated virus has been treated with nonionic detergent MESK with subsequent removal of viral cores by centrifugation. Supernatant was sterilized by filtration through the nuclear filters and cleared from detergent by dialysis. Glycoproteins obtained have not contained contaminating cellular or core viral proteins or viral shell lipids. In the absence of detergent, glycoproteins have formed the peculiar mycelial complexes. Biological activity of glycoproteins was kept at high level. Glycoproteins output at isolation from different strains of influenza viruses A, B and Sendai virus varied from 75 to 98%. Immunogenetic study of the preparations obtained has demonstrated their capability to stimulate the formation of antibodies against both viral glycoproteins comparable with the capability of intact virus. The obtained level of immunity was enough to protect organism against homologous infection. Samples of glycoproteins obtained are up to standards for subunit vaccines, and the technique of their preparation is perspective as far as the production of vaccine preparations is concerned.

Purification and concentration of rabies virus by chromatography on chemically modified porous silicates.

Tissue culture rabies virus was purified by gel chromatography on chemically modified macroporous glasses (MPG). Modification of the MPG surface by egg or human albumin prevented rabies virions from interaction with the glass surface so that all purified virus was eluted from the column. Chemically modified MPG could be used on a preparative scale for the purification of both infectious and inactivated rabies virus, the immunogenicity of the latter having been preserved. Protein content of the purified virus fractions was up to 4 microgram/ml, of which virus protein represented up to 5%.

[Chromatography of formalin-inactivated tick-borne encephalitis viruses on macroporous glass]. / Khromatografiia inaktivirovannogo formalinom virusa kleshchevogo éntsefalita na makroporistykh steklakh.

The results of further development of methods for chromatographic concentration and purification of tick-borne encephalitis virus on columns with porous glass using formalin-inactivated virus suspensions are presented. The method of adsorption chromatography on porous glass under optimal conditions concentrates inactivated TBE virus 20-40-fold with simultaneous purification from protein by 93%-95%. Inactivated virus was shown to keep on glass better than infectious virus. Gel filtration chromatography removes 99.2%-99.8% of protein impurities from inactivated TBE virus preparations giving a complete or nearly complete "yield" of virus particles. The sequential use of adsorption chromatography and gel filtration produced concentrated, most highly purified TBE virus preparations containing no more than 2 microgram/ml protein. Chromatographic virus preparations were immunologically active in experiments in laboratory animals.