Comparison of the Production of Recombinant Protein in Suspension Culture of CHO Cells in Spinner Flask and Shake Flask System

Abstract

Chinese hamster ovary (CHO) cells have been most widely used as the production host for the commercial production of biopharmaceuticals product. They have been extensively studied and developed, and today provide a stable platform for producing monoclonal antibodies and recombinant proteins. This study was focusing on comparison of suspension culture system by using spinner flask and shake flask for the growth and production of recombinant protein in CHO cell line. The CHO cells were transfected with an expression of DNA plasmid containing lac Z gene which codes for β-galactosidase. The recombinant genes in these CHO cells and the β-galactosidase expressing cells were adapted to suspension culture. The agitation speed for both spinner and shake flask were adjusted accordingly. The experiments were carried out in duplicate and samples were taken for cell count, determination of glucose consumption, lactate production and protein level by using biochemical assay. The result showed that, the cell growth in spinner flask is more favorable then in shake flask. The cell concentration in spinner flask is 58% higher than in shake flask. On the other hand, specific activity of β-galactosidase is 25% higher in spinner flask compared to shake flask, at the same agitation speed.

ABSTRAK: Sel ovari hamster China (Chinese hamster ovary (CHO)) digunakan secara meluas dalam hos pembiakan untuk tujuan komersil produk biofarmaseutikal. Ia telah dikaji dan dibangunkan secara ekstensif, dan kini ia menyediakan landasan yang stabil untuk penghasilan antibodi monoklon dan protein rekombinan. Kajian ini memfokuskan tentang penghasilan protein rekombinan menggunakan kultur ampaian sel CHO di dalam kelalang putar dan kelalang goncang. Sel CHO dimasukkan dengan plasmid DNA yang mengandungi gen lac Z yang juga memberikan kod untuk β-galaktosidase. Sel CHO β-galaktosidase-terungkap dimasukkan ke dalam kultur ampaian. Kelajuan agitasi untuk kedua-dua kelalang putar dan kelalang goncang disesuaikan dengan sewajarnya. Eksperimen dijalankan menggunakan pendua dan sampel yang diambil untuk kiraan sel, penentuan penggunaan glukosa, penghasilan laktat dan aras protein dengan menggunakan cerakin biokimia. Keputusan menunjukkan tumbesaran sel di dalam kelalang putar lebih menggalakkan daripada dalam kelalang goncang. Kepekatan sel dalam kelalang putar adalah 58% lebih tinggi daripada dalam kelalang goncang. Sebaliknya, pada kelajuan agitasi yang sama, aktiviti tertentu β-galaktosidase adalah 25% lebih tinggi dalam kelalang putar dibandingkan dengan kelalang goncang.