SU1109433A1 - Strain of hybrid cam cells used for preparing monoclonic antibodies for tick-borne encephalitis virus - Google Patents

Abstract

KEMA HYBRID CELL STRAIN USED TO OBTAIN MONOCLONAL ANTIBODIES TO THE TICK-BORN ENCEPHALITIS VIRUS.

The strain CEMA hybrid cells, No. 45/5 / (Collection of transplanted cell lines of the DI Ivanovsky Institute of Virology), used to produce monoclonal antibodies to tick-borne encephalitis virus. (L

Description

co oo oo The invention relates to medicine and concerns a new cell strain used to produce antibodies to tick-borne encephalitis virus. A cell strain that produces monoclonal antibodies to flavus viruses Dengue l is known. A known cell strain that produces monoclonal antibodies to the flare virus; it’s encephalitis 2. However, monoclonal antibodies of the first strain are not capable of reacting with tick-borne encephalitis virus, and antibodies produced by the second strain react with the non-pathogenic MOM Scalitz and have only anthma-hemagglutinating activity. The aim of the invention is to obtain monoclonal antibodies to the group for viruses of tick-borne encephalitis determinant. The goal is achieved by creating a strain of CEMA hybrid cells used to produce monoclonal antibodies against tick-borne encephalitis virus. The strain of transplantable CEMA myepia cells (hybridoma) was obtained by sludge of X63-Ag 8/653 mouse myeloma with spleen cells of T: encephalus, immunized with tick-borne encephalitis virus, and is a MYEKLONON hybrid of mouse cells. The strain of CEMA cells is stored in a collection of transplantable cell lines of the D.I. Institute of Virology. Ivano whom the Academy of Medical Sciences of the USSR under the number 45/5 / and is characterized by the following features. Cultural characteristics. The culture medium is ere da Dalbecco with 15% fetal bovine serum, sodium pyruvate 4 g / l. Glucose, antibiotic gates with cyn 50 µg / ml. Growth pattern stationary suspension. Shaking off method. Passaging frequency after 2-4 days. Sown dose of 150 00 cells in 1 ml. The multiplicity of sieving 1: 3,1: 5. Cell preservation. KEMA cells are frozen at 8, 15, AO passages. The total number of ampoules 50 to 5-10 million cells in the ampoule. The cryoprotective medium is a growth medium with 50% fetal calf serum and 10% glycerol. Survival rate when defrosting 73%. 32 Line contaminant information. No bacteria were detected, fungi and nuclei were not detected, mycoplasmas were not detected, viruses — particles of type A and C were found, similar to particles of the parent and Tamma X63-Ag8 / 653 cells. Isoenzymes, mobility of glscose-6-phosphate dehydrogenase and lactate dehydrogenase in cells of the CEMA clone is characteristic of murine cells. Culture morphologists. The culture consists of slightly attached round-shaped cells of varying size to the substrate, with dramas occupying most of the cell. The nuclei are large, there are duplex cells. Cytoplasm usually has a small rim or nucleus offset to one side of the cell. Caryology culture. (ariotype corresponds to the form (microscopic) With the C-staining method, stable markers were not detected. Modal class - 93 chromosomes (modal class for the parental myeloma line X63-Ag8 / 653 - 51 chromosome). Monoclonal antibody activity in serological reactions. Samples CEMA cells-induced ascites fluid (AF) and culture fluid (QL) are examined in the reaction of direct immunofluorescence (HMFA) and complement fixation (RBC) reactions. The reactions are performed using individual methods. Radiation of monoclonal antibodies in HMFA is given o in Table 1. The results of the experiments listed in Table 1 show that all samples M (and UC contain antibodies specific to tick-borne encephalitis virus. Control studies with VAL and Dengue 11 viruses are negative. The study of monoclonal antibodies to tick-borne encephalitis viruses in RSC is given in Table 2 B. The results of RAC show that monoclonal antibodies specifically react with tick-borne encephalitis antigen do not interact with antigens of VEL (alphavirus) and Deige 11 (ai virus) antigens. The use of the CEMA hybrid cell strain makes it possible to produce monocular, highly active specific antibodies to vysokopatogennomv

3t109A33

for a person, the tick-borne encephalitis virus strain eliminates the danger of contact with the virus, allows for improved methods for indicating the causative agent and diagnosing the disease, as well as obtaining and purifying the virus and viral drugs for the virus

tick-borne encephalitis.

Table 1

table 2

Claims (1)

CAM HYBRID CELL STRAIN USED TO OBTAIN MONOCLONAL ANTIBODIES TO TICK ESCEHALITIS VIRUS. KEMA hybrid cell strain, No. 45/5 / (Collection of transplantable cell lines of the D.I. Ivanovsky Institute of Virology), used to obtain monoclonal antibodies to tick-borne encephalitis virus. i109433 ΐ