References

vavilov

Вавиловский журнал генетики и селекции

Vavilov Journal of Genetics and Breeding

2500-3259

Institute of Cytology and Genetics of Siberian Branch of the RAS

10.18699/VJ19.490

vavilov-2016

Research Article

Молекулярная генетика

Molecular genetics

Влияние различий в третьем домене гликопротеина Е вируса клещевого энцефалита дальневосточного, сибирского и европейского субтипов на связывание рекомбинантных белков D3 с химерным антителом

The effect of differences in the third domain of the glycoprotein E of tick-borne encephalitis virus of the Far Eastern, Siberian and European subtypes on the binding of recombinant D3 proteins with a chimeric antibody

Байков

И. К.

Baykov

I. K.

ivan_baykov@mail.ru

Матвеев

А. Л.

Matveev

A. L.

Емельянова

Л. А.

Emelianova

L. A.

Каверина

Г. Б.

Kaverina

G. B.

Ткачёв

С. Е.

Tkachev

S. E.

Тикунова

Н. В.

Tikunova

N. V.

tikunova@niboch.nsc.ru

Институт химической биологии и фундаментальной медицины Сибирского отделения Российской академии наукРоссияInstitute of Сhemical Biology аnd Fundamental Medicine, SB RASRussian Federation

Институт химической биологии и фундаментальной медицины Сибирского отделения Российской академии наук; Новосибирский национальный исследовательский государственный университетРоссияInstitute of Сhemical Biology аnd Fundamental Medicine, SB RAS; Novosibirsk State UniversityRussian Federation

2019

14052019

233256261

2019

Байков И.К., Матвеев А.Л., Емельянова Л.А., Каверина Г.Б., Ткачёв С.Е., Тикунова Н.В.

Baykov I.K., Matveev A.L., Emelianova L.A., Kaverina G.B., Tkachev S.E., Tikunova N.V.

This work is licensed under a Creative Commons Attribution 4.0 License.

https://vavilov.elpub.ru/jour/article/view/2016

Целью настоящей работы было исследование влияния различий аминокислотной последовательности рекомбинантных доменов D3 гликопротеина Е вируса клещевого энцефалита дальневосточного, Сибирского и европейского субтипов на связывание протективного антитела ch14D5 с этими белками. Методами вестерн-блот анализа и поверхностного плазмонного резонанса было показано, что наибольшее сродство (KD= 1.7±0.5 нМ) антитело ch14D5 проявляет к домену D3 вируса клещевого энцефалита штамма «СофьинRu», принадлежащего к дальневосточному субтипу вируса. В то же время сродство к аналогичным белкам D3, полученным на основе штаммов «Заусаев», «1528-99» и «Абсеттаров» сибирского и европейского субтипов вируса клещевого энцефалита, оказалось заметно ниже (KD= 25±4, 300±50 и 250±50 нМ соответственно). Кроме того, информация о пространственном расположении аминокислотных остатков, которыми отличаются полученные рекомбинантные белки, указывает на то, что узнаваемый антителом ch14D5 эпитоп находится в области бокового ребра домена D3 гликопротеина Е.

Currently, a therapeutic drug based on recombinant antibodies for the prevention and treatment of tick-borne encephalitis virus (TBEV) is developed in ICBFM SB RAS, and the chimeric antibody ch14D5 is considered as one of the key components of this drug. It was previously shown that this antibody is directed to the domain D3 of the glycoprotein E of TBEV. It was previously shown that this antibody is able to protect mice from the European subtype of TBEV, strain “Absettarov”, and the presence of virus-neutralizing activity against the Far Eastern subtype of TBEV, strain 205 was also shown for this antibody. However, it remains unclear whether this antibody exhibits selectivity for different subtypes of TBEV. The aim of this study was to investigate the effect of amino acid sequence differences of recombinant D3 domains derived from the glycoprotein E of TBEV of the Far Eastern, Siberian and European subtypes on the binding of the protective antibody ch14D5 to these proteins. Using Western blot analysis and surface plasmon resonance, it was shown that ch14D5 antibody has the highest affinity (KD= 1.7±0.5 nM) for the D3 domain of the TBEV of the “Sofjin-Ru” strain belonging to the Far Eastern subtype of the virus. At the same time, the affinity of ch14D5 antibody for similar D3 proteins derived from “Zausaev”, “1528-99” and “Absettarov” strains of the Siberian and European subtypes of TBEV was noticeably lower (KD= 25±4, 300±50, 250±50 nM, respectively). In addition, information about the spatial arrangement of amino acid residues that are different for the studied recombinant proteins indicates that the epitope recognized by the ch14D5 antibody is in close proximity to the lateral ridge of D3 domain of E glycoprotein.

вирус клещевого энцефалитагликопротеин Едомен D3антителорекомбинантный белокповерхностный плазмонный резонанскартирование эпитопа

tick-borne encephalitis virusglycoprotein Edomain D3antibodyrecombinant proteinsurface plasmon resonanceepitope mapping

This work was supported by the Russian Federation Ministry of Science and Higher Education, RF President’s grant MK-6575.2018.4. The development of the protocol for obtaining recombinant D3 domains was supported by the Russian Science Foundation, project 17-74-10146. The design of the recomibinant D3_Sof domain of the Sof’in-Ru TBEV strain was supported by the same project. The production and purification of chimeric ch14D5 antibody was supported by the Russian Science Foundation, project 16-14-00083. The production of viral RNA and cDNA was supported by the Russian Foundation for Basic Research, project 18-44-540021.

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The authors declare that there are no conflicts of interest present.