Studies on the influence of different designs of eukaryotic vectors on the expression of recombinant IgA

Abstract Production and application of therapeutic monoclonal antibodies are second only to vaccines in the world pharmaceutical market. The most common therapeutic antibodies are monoclonal antibodies (mAbs) of the IgG isotype that are produced in eukaryotic CHO cells. In recent years, there has been a considerable interest in developing treatment medications based on IgA antibodies, which can have a wide range of effector functions on human mucous membranes. To study the expression level of immunoglobulin A (IgA) in mammal cells, we designed a set of bipromoter (CMV and EF1α) vectors. The vectors contain gene fragments that encode the heavy chain variable domain (VH) and the light chain variable domain (VL) of the human monoclonal antibody FI6v3 against the hemagglutinin of influenza virus A. They also contain gene fragments that encode the light chain (kappa type) constant domain and the heavy chain constant domain of the human antibody IgA1. The expression vectors differed in the orientation of the promoters and the presence or absence of introns. Two variants of the full-length light and heavy chains were cloned into a eukaryotic expression vector in head-to-head and head-to-tail orientations. The resulting plasmids were transfected into CHO-DG44 and HEK-293T cells. The antibody expression level for the stable transfection of CHO-DG44 and HEK-293T cell cultures was determined by ELISA. The results of the experiments showed that the expression of FI6v3-IgA1 antibodies significantly increased when eukaryotic cells were transfected with the plasmid pBiPr-ABIgA1FI6-Iht in which the heavy chain of IgA1 contains introns and the promoters are arranged head-to-tail..

Medienart:	E-Artikel

Erscheinungsjahr:	2017
Erschienen:	2017

Enthalten in:	Zur Gesamtaufnahme - volume:72
Enthalten in:	Moscow University biological sciences bulletin - 72(2017), 2 vom: Apr., Seite 63-68

Sprache:	Englisch

Beteiligte Personen:	Argentova, V. V. [VerfasserIn] Aliev, T. K. [VerfasserIn] Toporova, V. A. [VerfasserIn] Rybchenko, V. S. [VerfasserIn] Dolgikh, D. A. [VerfasserIn] Kirpichnikov, M. P. [VerfasserIn]

Links:	Volltext [lizenzpflichtig]

Themen:	Bi-promoter vector Eukaryotic expression Influenza A virus Intron Monoclonal antibody IgA1 Promoters of different orientation

doi:	10.3103/S0096392517020018

funding:
Förderinstitution / Projekttitel:

PPN (Katalog-ID):	SPR023273550