N- Method for preparing recombinant glycoproteins with improved N-glycan antennary structure by inhibiting biosynthesis of polylactosamine
In the present invention, it was confirmed that polylactosamine was reduced in recombinant human-derived erythropoietin produced by inhibiting (knocking down) the expression of the β3gnt2 gene, which is involved in polylactosamine biosynthesis, or from a cell line with the knocked out β3gnt2 gene, a...
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Zusammenfassung: | In the present invention, it was confirmed that polylactosamine was reduced in recombinant human-derived erythropoietin produced by inhibiting (knocking down) the expression of the β3gnt2 gene, which is involved in polylactosamine biosynthesis, or from a cell line with the knocked out β3gnt2 gene, and thus triple and/or quadruple antenna structures of N-glycan increased, leading to positive roles in increasing the half-life of glycoprotein and the efficacy of glycoprotein. Therefore, it was confirmed that a recombinant glycoprotein with an increased half-life can be produced through the method of the present invention.
본 발명은 폴리락토사민의 생합성에 관여하는 β3gnt2 유전자의 발현을 억제(녹다운)하거나 β3gnt2 유전자를 녹아웃시킨 세포주로부터 생산된 재조합 인간 유래 에리스로포이에틴에서 폴리락토사민의 감소를 확인, 그에 따른 N-글리칸의 삼중 및/또는 사중 안테나 구조 증가를 확인함으로써 당단백질의 반감기 증가 및 당단백질 효능에 긍정적인 역할이 가능함을 확인한 바, 본 발명의 방법을 통해 반감기를 증가시킨 재조합 당단백질의 제조가 가능함을 확인하였다. |
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