МОНОКЛОНАЛЬНЫЕ АНТИТЕЛА К ВИРУСУ БЕШЕНСТВА
Abstract
Hybridomas secreting monoclonal antibodies (MAbs) against rabies virus (RV) were prepared by hybridization
of splenocytes of BALB/c mice immunized by RV vaccine strain (ARRIAH strain) with the mouse
myeloma NSO/1 cell line. Properties of prepared MAbs were examined with indirect immunofluorescence
assay and solid-phase sandwich ELISA using different rabies virus strains and field isolates as well as with
rabies virus neutralization test in BHK-21 cell culture. As a result, three clones producing glycoprotein-specific
MAbs and one clone secreting Mabs to RV ribonucloeprotein (ARRIAH/N-1) were selected. RV glycoprotein-
specific ARRIAH/G-1 MAbs possessed virus-neutralizing activity and reacted with RV vaccine
strains only. Immunofluorescence assay with ARRIAH/N-1 MAbs detected RV ribonucleoprotein accumulations
in brain cells of animals died of rabies. ARRIAH/N-1 MAbs reacted with different RV strains and
field isolates.
of splenocytes of BALB/c mice immunized by RV vaccine strain (ARRIAH strain) with the mouse
myeloma NSO/1 cell line. Properties of prepared MAbs were examined with indirect immunofluorescence
assay and solid-phase sandwich ELISA using different rabies virus strains and field isolates as well as with
rabies virus neutralization test in BHK-21 cell culture. As a result, three clones producing glycoprotein-specific
MAbs and one clone secreting Mabs to RV ribonucloeprotein (ARRIAH/N-1) were selected. RV glycoprotein-
specific ARRIAH/G-1 MAbs possessed virus-neutralizing activity and reacted with RV vaccine
strains only. Immunofluorescence assay with ARRIAH/N-1 MAbs detected RV ribonucleoprotein accumulations
in brain cells of animals died of rabies. ARRIAH/N-1 MAbs reacted with different RV strains and
field isolates.
About the Authors
А. МолодкинRussian Federation
С. Аббасова
Филиал института биоорганической химии РАН, г. Пущино Московской
области
Russian Federation
Russian Federation
Н. Назаров
Russian Federation
Н. Балохина
Филиал института биоорганической химии РАН, г. Пущино Московской
области
Russian Federation
Russian Federation
В. Васильева
Филиал института биоорганической химии РАН, г. Пущино Московской
области
Russian Federation
Russian Federation
Х. Бозиев
Филиал института биоорганической химии РАН, г. Пущино Московской
области
Russian Federation
Russian Federation
Ф. Бровко
Филиал института биоорганической химии РАН, г. Пущино Московской
области
Russian Federation
Russian Federation
К. Груздев
Russian Federation
С. Рыбаков
Russian Federation
References
1. С.В. Грибенча, О.В. Василенко, В.А. Кузьмицкая и др. Взаимодействие моноклональных антител к структурным белкам вакцинного вируса Внуково-32 с другими вирусами группы бешенства // Вопр. вирусол. 1991. № 5. С. 399-402.
2. Н.А. Назаров, А.Е. Метлин, С.С. Рыбаков. Сравнительная характеристика антител, полученных на разные антигены вируса бешенства // Биол.- экол. пробл. заразных бол. диких ж-ных и их роль в патологии с.-х. ж-ных и людей: матер. Междунар. науч.-практ.-конф. окров, 2002. С. 168-170.
3. В.В. Недосеков, И.Ф. Вишняков, И.В. Никишин. Получение и использование иммуноасцитической жидкости, специфичной к вирусу бешенства // Пробл. инфекц. патологии с.-х. ж-ных: тез. докл. конф. Владимир, 1997. С. 182-183.
4. С.В. Грибенча, О.В. Василенко, В.А. Фуралев и др. Получение и характеристика гибридом, продуцирующих моноклональные антитела к струк- турным белкам вируса бешенства, штамм Внуково-32 // Вопр. вирусол. 1991. № 4. С. 318-321.
5. В.В. Недосеков, А.А. Стрижаков, А.Д. Середа,
6. Т.Р. Кирюхина. Получение и характеристика моноклональных антител к структурным белкам вируса бешенства штамм ТС-80 // Доклады РАСХН. 1999. № 4. С. 38-40.
7. А.П. Пономарев, Н.А. Назаров, С.С. Рыбаков и др. Совершенствование метода очистки и концентрирования вируса бешенства // Нейроинфекции: бешенство, губкообразная энцефалопатия КРС, Крейтцфельдта-Якоба и др. прионные болезни; листериоз, болезнь Ауески, болезнь Тешена: матер.Междунар.конф. Покров, 2001. С.49-50
8. J.D. Beatty, B.G. Beatty, W.G. Vlahos. Measurement of monoclonal antibody affinity by non-competitive enzyme immunoassay // J. Immunol. Methods. 1987. V. 100. P. 173-179.
9. P.L. Ey, S.J. Prowse, C.R. Leukin. Isolation of pure IgG1, IgG2a and IgG2b immunoglobulins from mouse serum using protein A-Sepharose // Mol. Immunol. 1978. V. 15. P. 429-436.
10. G. Kцhler, C. Milstein. Continuos cultures of fused cell secreting antibody of predefined specificity // Nature. 1975. V. 256. P. 495-497.
11. U. K. Laemlii. Cleavage of structural proteins during the assembly of the head of bacteriophage T4 // Nature. 1970. V. 227, N 5259. P. 680-685.
12. A.E. Metlin, J. Cox, S.S. Rybacov et al. Monoclonal antibody characterization of rabies virus isolates from Russia, Finland and Estonia // J. Vet. Med. 2004. V. 51. P. 94-96.
13. W.E. Marissen, R.A. Kramer, A. Rice et al. Novel rabies virus-neutralizing epitope recognized by human monoclonal antibody: fine mapping and escape mutant analysis // J. Virol. 2005. V. 79, N 8. P. 4672-4678.
14. P. Perrin. Techniques for the preparation of rabies conjugates // Laboratory techniques in rabies. 4th ed. Geneva, 1996. P. 433-444.
15. O.H. Lowry, N.J. Rosebrough, A.L. Farr, R.J. Randall. Protein measurement with Folin phenol reagent // J. Biol. Chemistry. 1951. V. 193. P. 265-275.
16. J.S. Smith, A.A. King. Monoclonal antibodies for the identification of rabies and non-rabies lyssaviruses // Laboratory techniques in rabies. - 4th ed. - Geneva, 1996. - P. 145-156.
17. C.V. Trimarchi, R.D. Rudd, M. Safford. An vitro virus neutralization test for rabies antibodi // Laboratory techniques in rabies. 4th ed. Geneva, 1996. P. 145-156.
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