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Transient expression in tobacco leaves of an aglycosylated recombinant antibody against the epidermal growth factor receptor
Author(s) -
Rodríguez Meilyn,
Ramírez Nadia I.,
Ayala Marta,
Freyre Freya,
Pérez Lincidio,
Triguero Ada,
Mateo Cristina,
SelmanHousein Guillermo,
Gavilondo Jorge V.,
Pujol Merardo
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20333
Subject(s) - recombinant dna , antibody , agrobacterium tumefaciens , biology , western blot , epidermal growth factor receptor , transgene , microbiology and biotechnology , immunofluorescence , agroinfiltration , receptor , gene expression , gene , biochemistry , immunology
When generating stably transformed transgenic plants, transient gene expression experiments are especially useful to rapidly confirm that the foreign molecule of interest is correctly assembled and retains its biological activity. TheraCIM® (CIMAB S.A., Havana) is a recombinant humanized antibody against the Epidermal Growth Factor receptor (EGF‐R), now in clinical trials for cancer therapy in Cuba and other countries. An aglycosylated version (Asn 297 was mutated for Gln 297) of this antibody was transiently expressed in tobacco leaves after vacuum‐mediated infiltration of recombinant Agrobacterium tumefaciens that carried a binary plasmid bearing the antibody heavy and light chain genes and plant regulation signals. Protein extracts from “agroinfiltrated” leaves were tested by ELISA and Western blot, showing that the fully assembled antibody was accumulated in plant tissues. The absence of plant specific glycans did not interfere in the assembling or in the activity of the plantibody, as demonstrated in this work. Indirect immunofluorescence demonstrated that the aglycosylated antibody expressed in plants recognizes the EGF‐R expressed on the surface of A431 human tumor culture cells. © 2004 Wiley Periodicals, Inc.