公开(公告)号：AT454444T

公开(公告)日：2010-01-15

申请号：AT04768294

申请日：2004-09-02

Applicant: UCB PHARMA SA

Inventor： GLOVER DAVID JOHN ,  SEHDEV MUKESH ,  REEKS DOMINIC GAMBIER

IPC: C12N1/21 ,  C07K16/00 ,  C12N1/20 ,  C12N15/72 ,  C12P21/02

Abstract: The present invention provides a method for controlling the partitioning of a recombinant protein between the supernatant and the periplasm in E. coli host cell cultures wherein expression of the recombinant protein by said cells is under the control of an inducible system, which method comprises: a) providing an E. coli host cell culture b) changing the growth rate of the E. coli host cells c) inducing expression of the recombinant protein wherein steps (b) and (c) can be performed in any order or simultaneously; and subsequently d) determining the yield of recombinant protein in the culture supernatant and the E. coli host cell periplasm e) comparing the yield determined in step (d) with the yield determined when at least one other growth rate has been used in step (b) f) selecting a growth rate from the comparison made in step (e) in which the partitioning of the recombinant protein between the supernatant and the periplasm is most suited to the primary recovery of the recombinant protein.

公开(公告)号：ES2339352T3

公开(公告)日：2010-05-19

申请号：ES04768294

申请日：2004-09-02

Applicant: UCB PHARMA SA

Inventor： GLOVER DAVID JOHN ,  SEHDEV MUKESH ,  REEKS DOMINIC GAMBIER

IPC: C12N1/21 ,  C07K16/00 ,  C12N1/20 ,  C12N15/72 ,  C12P21/02

Abstract: Un método para seleccionar una tasa de crecimiento para controlar la distribución de una proteína recombinante entre el sobrenadante y el periplasma en cultivos de células hospedantes de E. coli de modo que la distribución de la proteína recombinante sea más adecuada para la recuperación primaria de la proteína recombinante, en la que la expresión de la proteína recombinante por dichas células está bajo el control de un sistema inducible, cuyo método comprende: a) proporcionar un cultivo de células hospedantes de E. coli b) cambiar la tasa de crecimiento de las células hospedantes de E. coli tal que la tasa de crecimiento esté en el intervalo de 0,0005 a 0,04/h. c) inducir la expresión de la proteína recombinante en la que las etapas (b) y (c) pueden ser realizadas en cualquier orden o simultáneamente; y subsiguientemente d) determinar el rendimiento de proteína recombinante en el sobrenadante de cultivo y el periplasma de las células hospedantes de E. coli e) comparar el rendimiento determinado en la etapa (d) con el rendimiento determinado cuando al menos una otra tasa de crecimiento ha sido usada en la etapa (b) f) seleccionar una tasa de crecimiento a partir de la comparación hecha en la etapa (e) en la que la distribución de la proteína recombinante entre el sobrenadante y el periplasma es más adecuado para la recuperación primaria de la proteína recombinante.

公开(公告)号：PL1664278T3

公开(公告)日：2010-06-30

申请号：PL04768294

申请日：2004-09-02

Applicant: UCB PHARMA SA

Inventor： GLOVER DAVID JOHN ,  SEHDEV MUKESH ,  REEKS DOMINIC GAMBIER

IPC: C12N1/21 ,  C07K16/00 ,  C12N1/20 ,  C12N15/72 ,  C12P21/02

公开(公告)号：AU2004270929B2

公开(公告)日：2010-01-21

申请号：AU2004270929

申请日：2004-09-02

Applicant: UCB PHARMA SA

Inventor： REEKS DOMINIC GAMBIER ,  SEHDEV MUKESH ,  GLOVER DAVID JOHN

IPC: C12N1/20 ,  C12N1/21 ,  C12P21/02

Abstract: The present invention provides a method for controlling the partitioning of a recombinant protein between the supernatant and the periplasm in E. coli host cell cultures wherein expression of the recombinant protein by said cells is under the control of an inducible system, which method comprises: a) providing an E. coli host cell culture b) changing the growth rate of the E. coli host cells c) inducing expression of the recombinant protein wherein steps (b) and (c) can be performed in any order or simultaneously; and subsequently d) determining the yield of recombinant protein in the culture supernatant and the E. coli host cell periplasm e) comparing the yield determined in step (d) with the yield determined when at least one other growth rate has been used in step (b) f) selecting a growth rate from the comparison made in step (e) in which the partitioning of the recombinant protein between the supernatant and the periplasm is most suited to the primary recovery of the recombinant protein.

公开(公告)号：SG145696A1

公开(公告)日：2008-09-29

申请号：SG2008056434

申请日：2004-09-02

Applicant: UCB PHARMA SA

Inventor： GLOVER DAVID JOHN ,  SEHDEV MUKESH ,  REEKS DOMINIC GAMBIER

IPC: C12N1/20 ,  C12N1/21 ,  C12P21/02

Abstract: METHODS FOR PRODUCING RECOMBINANT PROTEINS The present invention provides a method for controlling the partitioning of a recombinant protein between the supernatant and the periplasm in E.coli host cell cultures wherein expression of the recombinant protein by said cells is under the control of an inducible system, which method comprises: a) providing an E.coli host cell culture b) changing the growth rate of the E.coli host cells c) inducing expression of the recombinant protein wherein steps (b) and (c) can be performed in any order or simultaneously; and subsequently d) determining the yield of recombinant protein in the culture supernatant and the E.coli host cell periplasm e) comparing the yield determined in step (d) with the yield determined when at least one other growth rate has been used in step (b) f) selecting a growth rate from the comparison made in step (e) in which the partitioning of the recombinant protein between the supernatant and the periplasm is most suited to the primary recovery of the recombinant protein.