Vector and mammalian cell line having improved productivity

Details Vector and mammalian cell line having improved productivity Vector and mammalian cell line having improved productivity

C - Chemistry, Metallurgy

12

N

C12N 15/85 (2006.01) C07K 14/755 (2006.01) C12N 15/57 (2006.01) C12N 15/69 (2006.01) C12Q 1/04 (2006.01)

Patent

CA 2125449

An improved vector for introducing a DNA coding sequence for a heterologous protein into a mammalian cell line to increase protein expression. The vector comprises a cDNA protein coding sequence, a first promoter and a downstream donor intron, the first promoter and downstream donor intron being located between a second promoter and a donor intron acceptor. In an illustrative example, the sequence codes for Factor VIII and high productivity of a transfected cell line is assured by establishing a minimum amplifiability index which, contrary to current practice, focuses on using cell lines having relatively low productivity prior to amplification.

L'invention concerne un vecteur amélioré permettant d'introduire une séquence codante d'ADN pour une protéine hétérologue dans une lignée cellulaire de mammifères pour accroître l'expression de protéine. Le vecteur comprend une séquence codante d'ADNc, un premier promoteur et un intron donneur aval situé entre un deuxième promoteur et un accepteur d'intron donneur. € titre d'exemple, on assure les codes de séquence pour le facteur VIII et la productivité élevée d'une lignée cellulaire transfectée en établissant un indice d'amplifiabilité minimal qui, contrairement à la pratique courante, met l'accent sur l'utilisation de lignées cellulaires ayant une productivité relativement faible avant l'amplification.

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