TY - JOUR
T1 - Efficient production of single-chain fragment variable-based N-terminal trimerbodies in Pichia pastoris
AU - Blanco-Toribio, Ana
AU - Lacadena, Javier
AU - Nuñez-Prado, Natalia
AU - Álvarez-Cienfuegos, Ana
AU - Villate, Maider
AU - Compte, Marta
AU - Sanz, Laura
AU - Blanco, Francisco J.
AU - Álvarez-Vallina, Luis
N1 - Funding Information:
This study was supported by grants from Ministerio de Economía y Competitividad (BIO2011–22738), and Comunidad de Madrid (S-BIO-0236– 2006 and S2010/BMD-2312) to L.A-V.; from Fondo de Investigación Sanitaria/ Instituto de Salud Carlos III (PI13/00090) to L.S.; and from Ministerio de Economía y Competitividad (BFU2012-32404) to J.L. A.B-T. was supported by Programa Torres Quevedo from Ministerio de Economía y Competitividad, cofounded by the European Social Fund (PTQ11–04604).
Publisher Copyright:
© 2014 Blanco-Toribio et al.
PY - 2014/8/12
Y1 - 2014/8/12
N2 - Background: Recombinant antibodies are highly successful in many different pathological conditions and currently enjoy overwhelming recognition of their potential. There are a wide variety of protein expression systems available, but almost all therapeutic antibodies are produced in mammalian cell lines, which mimic human glycosylation. The production of clinical-grade antibodies in mammalian cells is, however, extremely expensive. Compared to mammalian systems, protein production in yeast strains such as Pichia pastoris, is simpler, faster and usually results in higher yields.Results: In this work, a trivalent single-chain fragment variable (scFv)-based N-terminal trimerbody, specific for the human carcinoembryonic antigen (CEA), was expressed in human embryonic kidney 293 cells and in Pichia pastoris. Mammalian- and yeast-produced anti-CEA trimerbody molecules display similar functional and structural properties, yet, the yield of trimerbody expressed in P. pastoris is about 20-fold higher than in human cells.Conclusions: P. pastoris is an efficient expression system for multivalent trimerbody molecules, suitable for their commercial production.
AB - Background: Recombinant antibodies are highly successful in many different pathological conditions and currently enjoy overwhelming recognition of their potential. There are a wide variety of protein expression systems available, but almost all therapeutic antibodies are produced in mammalian cell lines, which mimic human glycosylation. The production of clinical-grade antibodies in mammalian cells is, however, extremely expensive. Compared to mammalian systems, protein production in yeast strains such as Pichia pastoris, is simpler, faster and usually results in higher yields.Results: In this work, a trivalent single-chain fragment variable (scFv)-based N-terminal trimerbody, specific for the human carcinoembryonic antigen (CEA), was expressed in human embryonic kidney 293 cells and in Pichia pastoris. Mammalian- and yeast-produced anti-CEA trimerbody molecules display similar functional and structural properties, yet, the yield of trimerbody expressed in P. pastoris is about 20-fold higher than in human cells.Conclusions: P. pastoris is an efficient expression system for multivalent trimerbody molecules, suitable for their commercial production.
KW - Monoclonal antibody
KW - Multivalent antibody
KW - P. Pastoris
KW - Recombinant antibody
KW - Trimerbody
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U2 - 10.1186/s12934-014-0116-1
DO - 10.1186/s12934-014-0116-1
M3 - Article
C2 - 25112455
AN - SCOPUS:84946410301
SN - 1475-2859
VL - 13
JO - Microbial Cell Factories
JF - Microbial Cell Factories
IS - 1
M1 - 116
ER -