New Genetic Constructs for Generation of Stable Therapeutic Antibodies to Organophosphorus Toxins in Methylotrophic Yeasts Pichia Pastoris
- Authors: Mokrushina Y.A.1, Stepanova A.V.1, Bobik T.V.1, Smirnov I.V.1, Gabibov A.G.1
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Affiliations:
- Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
- Issue: Vol 161, No 1 (2016)
- Pages: 83-87
- Section: Article
- URL: https://journals.rcsi.science/0007-4888/article/view/237123
- DOI: https://doi.org/10.1007/s10517-016-3351-0
- ID: 237123
Cite item
Abstract
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.
About the authors
Yu. A. Mokrushina
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Russian Federation, Moscow
A. V. Stepanova
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Russian Federation, Moscow
T. V. Bobik
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Russian Federation, Moscow
I. V. Smirnov
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Author for correspondence.
Email: smirnov@mx.ibch.ru
Russian Federation, Moscow
A. G. Gabibov
Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences
Email: smirnov@mx.ibch.ru
Russian Federation, Moscow