Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line

Evandra Strazza Rodrigues; Virgínia Picanço-Castro; Marta Regina Espanhol; Luiz Alberto Martins de Andrade; Patricia Vianna Bonini Palma; Simone Kashima; Aparecida Maria Fontes; Dimas Tadeu Covas

doi:10.1186/2193-1801-2-328

Quantitative correlation between transcriptional levels of ER chaperone, peroximal protein and FVIII productivity in human Hek-293 cell line

Springerplus. 2013 Jul 18;2(1):328. doi: 10.1186/2193-1801-2-328. eCollection 2013.

Authors

Evandra Strazza Rodrigues¹, Virgínia Picanço-Castro², Marta Regina Espanhol², Luiz Alberto Martins de Andrade², Patricia Vianna Bonini Palma², Simone Kashima¹, Aparecida Maria Fontes³, Dimas Tadeu Covas⁴

Affiliations

¹ Hemotherapy Center of Ribeirão Preto , Faculty of Medicine in Ribeirão Preto-FMRP, University of São Paulo-USP, São Paulo, Brazil ; Faculty of Pharmaceutical Sciences in Ribeirão Preto-FCFRP, University of São Paulo-USP, São Paulo, Brazil ; Hemocentro de Ribeirão Preto, National Institute of Science and Technology in Stem Cell and Cell Therapy, Avenida Tenente Catão Roxo, 2501 - Ribeirão Preto, São Paulo, 14051-140 Brazil.
² Hemotherapy Center of Ribeirão Preto , Faculty of Medicine in Ribeirão Preto-FMRP, University of São Paulo-USP, São Paulo, Brazil.
³ Hemotherapy Center of Ribeirão Preto , Faculty of Medicine in Ribeirão Preto-FMRP, University of São Paulo-USP, São Paulo, Brazil ; Faculty of Medicine in Ribeirão Preto-FMRP, University of São Paulo-USP, São Paulo, Brazil.
⁴ Hemotherapy Center of Ribeirão Preto , Faculty of Medicine in Ribeirão Preto-FMRP, University of São Paulo-USP, São Paulo, Brazil ; Faculty of Medicine in Ribeirão Preto-FMRP, University of São Paulo-USP, São Paulo, Brazil ; Hemocentro de Ribeirão Preto, National Institute of Science and Technology in Stem Cell and Cell Therapy, Avenida Tenente Catão Roxo, 2501 - Ribeirão Preto, São Paulo, 14051-140 Brazil.

Abstract

Hek-293 cell line presents good production platform for recombinant therapeutic proteins, however little is known about the components that contribute to the cellular control of recombinant protein production. In this study, we generated a Hek-293 producing recombinant factor VIII (FVIII) and we evaluated the immunoglobulin-binding protein (BiP) and phytanoil-CoA α-hydroxylase (PAHX) expression levels which are known for diminishing FVIII production. Our analyses showed that the recombinant cell population expresses 3.1 ± 1.4 fold of BIP mRNA (P = 0.0054) and 97.8 ± 0.5 fold of PAHX mRNA (P = 0.0016) compared to nontransduced cells. The amount of these proteins was inversely correlated to the secreted FVIII. In conclusion, BIP and PAHX expression are augmented in human cells producing FVIII and they antagonize the amount of therapeutic factor VIII in the cell culture.

Keywords: ER BIP chaperone; Factor VIII; Hemophilia A; PAHX gene expression and Recombinant Factor VIII∆B.