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

نویسندگان

  • 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
چکیده

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.

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

منابع مشابه

The Effects of Novel Mutations in A1 Domain of Human Coagulation Factor VIII on Its Secretion Level in Cultured Mammalian Cells

Inefficient secretion of the human coagulation factor (hFVIII) in mammalian expression systems is one ofthe main causes of the hFVIII low expression level, attributed to its interaction with a chaperone known asBiP/GRP78. In order to improve secretion efficiency of the hFVIII, based on the higher secretion level of theporcine FVIII and analysis of the hFVIII A110 region, that ...

متن کامل

Evaluation of Iranian Snake ‘Macrovipera lebetina’ Venom Cytotoxicity in Kidney Cell Line HEK-293

Background:Envenomation by Macrovipera lebetina (M. lebetina) is characterized by prominent local tissue damage, hemorrhage, abnormalities in the blood coagulation system, necrosis, and edema. However, the main cause of death after a bite by M. lebetina has been attributed to acute renal failure (ARF). It is unclear whether the venom components have a direct or indirect action in causing ARF. T...

متن کامل

P-157: Polymorphic Core Promoter GA-repeats Alter Gene Expression of The Early Embryonic Developmental Genes

Background: We examine the GA-repeat core promoters of MECOM and GABRA3 in human embryonic kidney-293 cell line and show that those GA-repeats have promoter activity,and those different alleles of the repeats can significantly alter gene expression.We propose a novel role for GA-repeat core promoters to regulate gene expression in the genes involved in development and evolution. Materials and M...

متن کامل

The F309S mutation increases factor VIII secretion in human cell line

OBJECTIVES The capacity of a human cell line to secrete recombinant factor VIII with a F309S point mutation was investigated, as was the effect of the addition of chemical chaperones (betaine and sodium-4-phenylbutyrate) on the secretion of factor VIII. METHODS This work used a vector with a F309S mutation in the A1 domain to investigate FVIII production in the HEK 293 human cell line. Factor...

متن کامل

تولید هورمون رشد انسانی نوترکیب توسط سلول تخمدان هامستر چینی و بررسی فعالیت زیستی آن به روش سنجش گزارشگر ژنی

 Background: Cultivated mammalian cells, because of their capacity for proper protein folding, assembly and post–translational modification, have become the dominant system for production of recombinant proteins in clinical application. Therefore, the quality and efficacy of protein can be superior when expressed in mammalian cells compared to other hosts such as bacteria. Gene reporte...

متن کامل

ذخیره در منابع من


  با ذخیره ی این منبع در منابع من، دسترسی به آن را برای استفاده های بعدی آسان تر کنید

برای دانلود متن کامل این مقاله و بیش از 32 میلیون مقاله دیگر ابتدا ثبت نام کنید

ثبت نام

اگر عضو سایت هستید لطفا وارد حساب کاربری خود شوید

عنوان ژورنال:

دوره 2  شماره 

صفحات  -

تاریخ انتشار 2013