Preview

Russian Journal of Biotherapy

Advanced search

Аffinity properties of plant-made anti-her2 antibodies

https://doi.org/10.17650/1726-9784-2018-17-1-95-100

Abstract

The Her2 receptor is an important target for antitumor therapy in the treatment of breast cancer. Trastuzumab, based on anti-Her2 monoclonal antibodies, is used in clinical practice. Trastuzumab is produced by animal cells culture technology and is quite expensive. We use the technology of production of recombinant antibodies in the plants Nicotiana benthamiana with a high yield of final purified protein. Objective. The aim of following study is a comparison of monoclonal antibodies received via classic cell culture technology and produced in plant biomass. Materials and methods. Recombinant plant-made antibodies were isolated by affinity chromatography from the biomass of N. benthamiana plants agroinfiltrated by vector constructs. Comparison of affinity properties was carried out by immunocytochemical staining of cells and competitive binding using flow cytometry analysis. Results and conclusion. We show that the antibodies expressed in N. benthamiana are equal to those obtained from mammalian cells in binding to Her2 antigen localized on the surface of the SK-BR-3 cells. In the present work it was shown that the plant-made anti-Her2 antibodies do not differ in specific binding with the Her2 antigen, as well as with IV subdomain of the Her2 receptor.

About the Authors

A. I. Scherbakov
N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia
Russian Federation
24 Kashirskoe Shosse, Moscow 115478


E. N. Kosobokova
N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia
Russian Federation
24 Kashirskoe Shosse, Moscow 115478


M. V. Pinyugina
N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia
Russian Federation
24 Kashirskoe Shosse, Moscow 115478


E. V. Sheshukova
N.I. Vavilov Institute of General Genetics, Russian Academy of Sciences
Russian Federation
3 Gubkina St., Moscow 119991


V. S. Kosorukov
N.N. Blokhin National Medical Research Center of Oncology, Ministry of Health of Russia
Russian Federation
24 Kashirskoe Shosse, Moscow 115478


References

1. Давыдов М.И. Структура заболеваемости злокачественными новообразованиями населения России в 2008 г. Вестник РОНЦ им. Н.Н. Блохина РАМН 2010;21(2):52.

2. Чиссов В.И. Злокачественные новообразования в России в 2010 году (заболеваемость и смертность). М.: ФГБУ «МНИОИ им. П.А. Герцена» Минздравсоцразвития России, 2012. 260 с.

3. Hudziak R.M., Lewis G.D., Winget M. et al. p185HER2 monoclonal antibody has antiproliferative effects in vitro and sensitizes human breast tumor cells to tumor necrosis factor. Mol Cell Biol 1989;3:1165–72. DOI: 10.1128/MCB.9.3.1165.

4. Komarova T.V., Kosorukov V.S., Frolova O.Y. et al. Plant-made trastuzumab (herceptin) inhibits HER2/Neu+ cell proliferation and retards tumor growth. PLoS One 2011:e17541. DOI: 10.1371/journal.pone.0017541.

5. Кособокова Е.Н., Пинюгина М.В., Косоруков В.С. Получение биологически активного интерферона-а-2b человека из растений Nicotiana benthamiana. Биотехнология 2015;4:52–61.

6. Steplewski Z., Sun L.K., Shearman C.W. et al. Biological activity of human – mouse IgG1, IgG2, IgG3, and IgG4 chimeric monoclonal antibodies with antitumor specificity. Proc Natl Acad Sci USA 1988;85(13):4852–6. PMID: 3387441.

7. Косоруков В.С., Скрыпник К.А., Трещалина Е.М., Андронова Н.В. Противоопухолевое действие субстанции рекомбинантных антител против онкоантигена HER2/neu, полученной из растительного продуцента. Российский биотерапевтический журнал 2012;11(2):4.

8. Dorokhov Y.L., Frolova O.Y., Skurat E.V. et al. A novel function for a ubiquitous plant enzyme pectin methylesterase: the enhancer of RNA silencing. FEBS Lett 2006;580:3872–8. DOI: 10.1016/j.febslet.2006.06.013.

9. Komarova T.V., Sheshukova E.V., Kosobokova E.N. et al. Trastuzumab and pertuzumab plant biosimilars: Modification of Asn297-linked glycan of the mAbs produced in a plant with fucosyltransferase and xylosyltransferase gene knockouts. Biochemistry (Moscow) 2017;82(4):510–20. DOI: 10.1134/S0006297917040137.

10. Косоруков В.С., Кособокова Е.Н., Пинюгина М.В. и др. Биологическая активность рекомбинантных антител против рецептора Her2, полученных из растительного источника. Российский биотерапевтический журнал 2015;14(2):105–12.


Review

For citations:


Scherbakov A.I., Kosobokova E.N., Pinyugina M.V., Sheshukova E.V., Kosorukov V.S. Аffinity properties of plant-made anti-her2 antibodies. Russian Journal of Biotherapy. 2018;17(1):95-100. (In Russ.) https://doi.org/10.17650/1726-9784-2018-17-1-95-100

Views: 576


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


ISSN 1726-9784 (Print)
ISSN 1726-9792 (Online)