Human hybrid host cell for mammalian gene expression

Details Human hybrid host cell for mammalian gene expression Human hybrid host cell for mammalian gene expression

C - Chemistry, Metallurgy

12

N

C12N 15/01 (2006.01) C12N 5/02 (2006.01) C12N 5/10 (2006.01) C12N 5/22 (2006.01) C12N 5/24 (2006.01) C12N 15/02 (2006.01) C12N 15/07 (2006.01) C12N 15/08 (2006.01) C12N 15/09 (2006.01) C12P 21/00 (2006.01)

Patent

CA 2353698

Human/human hybrid cells were made via fusion of human embryonic kidney cells (293S) and modified Burkitt's lymphoma cells (2B8). The fusion cells are useful as host cells for the recombinant expression of mammalian genes. The advantages of using these hybrid clones of human kidney- and B-cells, called HKBs, for mammalian gene expression, include (i) the cells are negative for immunoglobulin expression, (ii) the cells grow easily in plasma protein-free medium (with or without the addition of recombinant insulin) as suspension cultures in a shake flask or in a fermenter (iii) the cells are very susceptible for transfection of DNA, and (iv) the cells secrete high levels of heterologous recombinant proteins, such as recombinant monoclonal antibodies, soluble ICAM-1, rIL-4, and rFVIII.

L'invention concerne des cellules hybrides humaines/humaines réalisées par fusion de cellules de rein embryonnaires (293S) et de cellules modifiées du lymphome de Burkitt (2B8). Les cellules de fusion conviennent en tant que cellules hôtes pour l'expression recombinante de gènes de mammifères. Concernant l'expression génétique chez les mammifères, ces clones hybrides de rein humain et de lymphocytes B, appelés HKB, présentent les avantages suivants : (i) les cellules sont négatives pour l'expression des immunoglobulines, (ii) les cellules se multiplient aisément dans un milieu exempt de protéines plasmatiques (avec ou sans ajout d'insuline de recombinaison), en cultures en suspension dans un flacon incubé sous agitation ou dans un fermenteur (iii) les cellules se prêtent bien à la transfection par l'ADN et (iv) les cellules sécrètent à un niveau élevé des protéines de recombinaison hétérologues, telles que des anticorps monoclonaux de recombinaison, ICAM-1 soluble, rIL-4 et rFVIII.

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