In-process control in a method for producing epo

Details In-process control in a method for producing epo In-process control in a method for producing epo

C - Chemistry, Metallurgy

07

K

C07K 14/505 (2006.01) C07K 1/28 (2006.01) C12P 21/02 (2006.01) G01N 33/68 (2006.01) G01N 33/74 (2006.01) G01N 27/447 (2006.01) G01N 33/543 (2006.01)

Patent

CA 2747320

The invention relates to a method for determining the isoform composition of erythropoietin, comprising the following steps: a) isoelectrical focusing of a sample comprising erythropoietin in a gel over a pH range having a lower limit of 2.5 to 3.5 and an upper limit of 5 to 8, wherein the sample comprising erythropoietin originates from a culture supernatant of erythropoietin producing eukaryotic cells; b) transferring of the proteins comprised and separated in the gel to a membrane; c) verifying the erythropoietin bound to the membrane by specific antibodies; and to a method for in-process control of culture supernatants of erythropoietin producing eukaryotic cells during the fermentative production process.

L'invention concerne un procédé de détermination de la composition d'isoformes de l'érythropoïétine, comprenant les étapes suivantes : a) focalisation isoélectrique d'un échantillon contenant de l'érytropoïétine dans un gel, dans un domaine de pH dont la limite inférieure se situe entre 2,5 et 3,5 et dont la limite supérieure se situe entre 5 et 8, l'échantillon contenant l'érythropoïétine provenant d'un surnageant de culture de cellules eucaryotiques produisant de l'érythropoïétine; b) transfert, sur une membrane, de la protéine contenue dans le gel et séparée; c) caractérisation de l'érytropoïétine liée à la membrane, par des anticorps spécifiques; et mise en oeuvre d'un procédé de contrôle, lors de la fabrication, de surnageants de culture de cellules eucaryotiques produisant de l'érythropoïétine, dans le cadre d'un processus de production par fermentation.

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