New Genetic Constructs for Generation of Stable Therapeutic Antibodies to Organophosphorus Toxins in Methylotrophic Yeasts Pichia Pastoris
- Авторы: Mokrushina Y.1, Stepanova A.1, Bobik T.1, Smirnov I.1, Gabibov A.1
-
Учреждения:
- Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
- Выпуск: Том 161, № 1 (2016)
- Страницы: 83-87
- Раздел: Article
- URL: https://journals.rcsi.science/0007-4888/article/view/237123
- DOI: https://doi.org/10.1007/s10517-016-3351-0
- ID: 237123
Цитировать
Аннотация
We propose a new method of obtaining of stable Fab-fragments of antibodies in Pichia pastoris expression system. Recently, we obtained Fab-fragments of antibodies neutralizing organophosphorus toxins. However, high yield of the target products was not attained because of high level of proteolytic degradation. In the present study, we identified sites of proteolytic degradation in Fab-fragments and endogenous proteases performing degradation, which allowed obtaining optimized genetic constructs for expression of antibody heavy chains (IgGγ1) and kappa and lambda isotypes of light chains. Co-transformation of these vectors allowed obtaining Fab-fragments of antibodies to organophosphorus toxins without proteolytic degradation of the product.
Об авторах
Yu. Mokrushina
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Россия, Moscow
A. Stepanova
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Россия, Moscow
T. Bobik
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Россия, Moscow
I. Smirnov
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Автор, ответственный за переписку.
Email: smirnov@mx.ibch.ru
Россия, Moscow
A. Gabibov
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Россия, Moscow