Experimental Substantiation of Cultural Technologies Introduction into Manufacturing of Anti-Rabies Immunoglobulin

The review provides information on the major outcomes of research and development work, performed within the frames of the Federal Target Program “National system of chemical and biological safety” (2009-2014), aimed at elaboration and introduction of cultural techniques into the manufacturing of anti-rabies immunoglobulin. Described are the key phases in methodology engineering, deployed for the large-scale cultivation of fixed rabies virus, concentration of cultural liquid, quantitation of rabies virus using PCR, and immunization of producers. Obtained pilot batches of the enhanced anti-rabies immunoglobulin, complying with normative requirements to commercial formulations, testify to the effectiveness of the developed biotechnological and methodological procedures, as well as of the designed engineering-manufacturing project documentation

Optimization of Specifications for Scaled-Up Fixed Rabies Virus Cultivation (“Moscow 3253” Strain) in Vero Cell Culture

/cell, using maintenance media 199 with admixture of 0.1 % human serum albumin or 2 % bovine serum. Optimum media volume in the roll-bottle for fixed rabies virus strain, “Moscow 3253”, cultivation is 200-400 ml. It depends upon the proliferating surface area. Specifications stated above provide for the obtainment of culture liquid with rabies virus titer - 1:256 - 1:512, if assayed in ELISA. This cultural virus is recommended as a basis for immunization material obtainment with a view to produce anti-rabies immunoglobulin from equine blood serum

Cultural Antigen in the Technology for Anti-Rabies Immunoglobulin Obtainment from Equine Blood Serum

week after immunization (specific activity is identified using neutralization reaction on the model of white mice and dot-blot immunoassay). This level of activity is sufficient for the fractioning of immune serum and extraction of anti-rabies immunoglobulin. Physicochemical and biological properties of the anti-rabies immunoglobulin, obtained with the help of cultural antigen technique, meet the requirements stated in the normative documentation on anti-rabies immunoglobulins extracted from equine blood serum. Specific activity level of experimental batches of anti-rabies immunoglobulin, obtained with the help of cultural technologies, corresponds to 242 and 214 IU/ml

ISOLATION OF GLICOPROTEID FROM THE FIXED RABIES VIRUS, STRAIN «MOSCOW 3253», AND CONSTRUCTING OF DOT-IMMUNOASSAY DIAGNOSTICUM ON ITS BASIS

Described here are the results of glicoproteid isolation from the fixed rabies virus, strain «Moscow 3253», using non-ionic detergent with subsequent chromatographic purification. The obtained antigen was demonstrated to be applicable as immunoreagent for construction of diagnosticum, by means of conjugation with colloid gold nanoparticles. The diagnosticum is meant for detection of specific antibodies in immune sera of horsesproducers, and in the preparation of anti-rabies immunoglobulin, in dot-immunoassay

Production of Rabbit Anti-Rabies Immunoglobulin Using Cultural Antigen

Justified is the possibility of application of fixed rabies virus Moscow 3253, reproduced on the cell culture Vero, as an antigen for heterologous anti-rabies immunoglobulin production. Application of the adjuvant - aluminium hydrate - is determined to be effective. Herein, by the day 73 post immunization, specific antibodies titer is ≥ 1:500 (wherein specific activity has been identified by means of neutralization test, carried out on white mice, and dot-blot immunoassay). The level of specific activity in experimental samples of anti-rabies immunoglobulin, isolated from rabbit immune serum, corresponds to 332 and 347 ME/ml. Physicochemical and biological properties of anti-rabies immunoglobulin, produced with the help of cultural antigen, fully comply with regulatory requirements specified for commercial preparation of heterologous anti-rabies immunoglobulin