The F309S mutation increases factor VIII secretion in human cell line
Rev. bras. hematol. hemoter
; 38(2): 135-140, graf
Article
en En
| LILACS
| ID: lil-787663
Biblioteca responsable:
BR408.1
ABSTRACT
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 VIII activity was measured by chromogenic assay. Furthermore, the effects of chemical drugs on the culture were evaluated.RESULTS:
The addition of the F309S mutation to a previously described FVIII variant increased FVIII secretion by 4.5 fold. Moreover, the addition of betaine or sodium-4-phenylbutyrate increased the secretion rate of FVIIIÄB proteins in HEK 293 cells, but the same effect was not seen for FVIIIÄB-F309S indicating that all the recombinant protein produced had been efficiently secreted.CONCLUSION:
Bioengineering factor VIII expressed in human cells may lead to an efficient production of recombinant factor VIII and contribute toward low-cost coagulation factor replacement therapy for hemophilia A. FVIII-F309S produced in human cells can be effective in vivo.Palabras clave
Texto completo:
1
Colección:
01-internacional
Banco de datos:
LILACS
Asunto principal:
Fenilbutiratos
/
ADN Recombinante
Límite:
Humans
Idioma:
En
Revista:
Rev. bras. hematol. hemoter
Asunto de la revista:
HEMATOLOGIA
Año:
2016
Tipo del documento:
Article
País de afiliación:
Brasil