Genetic Engineering of Native Chain Combinations of B-Cell Repertoires on the Surface of Methylotrophic Yeasts Pichia pastoris


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Abstract

We designed genetic constructs for exposing Fab-fragment library of natively paired single cell B-cell receptors on the surface of Pichia pastoris yeast cells. We have previously obtained the A17 antibody in our laboratory [6]. In this study we showed that the newly designed genetic constructs provide a compatible level of A17 antibody Fab fragment on the surface of yeast cells as well as in the case of vectors containing DNA fragments corresponding to each chain of the antibody. The data suggest that the developed approach for constructing immunoglobulin gene libraries is adequate and fully convenient for studying properties of the real human B-lymphocyte repertoire.

About the authors

T. V. Bobik

Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Email: gabibov@mx.ibch.ru
Russian Federation, Moscow

E. M. Shurdova

Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Email: gabibov@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

Email: gabibov@mx.ibch.ru
Russian Federation, Moscow

N. A. Ponomarenko

Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Email: gabibov@mx.ibch.ru
Russian Federation, Moscow

E. N. Khurs

Laboratory of Chemical Bases and Biocatalysis V. A. Engelhardt Institute of Molecular Biology, Russian Academy of Sciences

Email: gabibov@mx.ibch.ru
Russian Federation, Moscow

V. D. Knorre

Laboratory of Biocatalysis, M. M. Shemyakin and Yu. A. Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences

Email: gabibov@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

Author for correspondence.
Email: gabibov@mx.ibch.ru
Russian Federation, Moscow

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