Transferin manusia (hTf) memainkan peranan yang penting dalam fungsi bakteriostatik dan pengangkutan ferum dari bahagian penyimpanan ke sel–sel yang membiak melalui proses endositosis janaan reseptor. Sistem ekspresi bakulovirus sel serangga telah dipakai secara meluas sebagai sistem alternatif dalam penghasilan Transferin manusia rekombinan (rhTf). Kajian ini ditumpukan ke atas pengoptimuman glutamina, glukosa dan campuran lipid 1000x yang dapat meningkatkan penghasilan rhTf. Reka bentuk eksperimen yang melibatkan 17 eksperimen reka bentuk komposit berpusat (CCD) telah digunakan dan hasil kajian dianalisis oleh Statistika (Statsoft v. 5.0). Metodologi permukaan tindak balas (RSM) telah mengenalpasti nilai optimum parameterparameter yang dikaji iaitu glutamina=2211.20 mg/L, glukosa=1291.95 mg/L, dan campuran lipid 1000x=0.64 %v/v. Hasil optimasi menunjukkan peningkatan hasil rhTf sebanyak tiga kali ganda, iaitu daripada 19.89 μg/ml kepada 65.12 μg/ml.
Kata kunci: Transferin manusia; bakulovirus sel serangga; reka bentuk eksperimen; reka bentuk komposit berpusat; metodologi permukaan tindak balas
Human Transferrin (hTf) plays a big role in providing bacteriostatic functions as well as to transport iron from the storage part to all proliferating cells by receptor mediated endocytosis. Insect cells baculovirus expression system has been widely used as an alternative expression system for the production of recombinant human Transferrin (rhTf). This work focused mainly on the optimization of glutamine, glucose and lipid mixtures 1000x to increase rhTf yield. An experimental design involving 17 central composite design (CCD) experiments was employed and results were analyzed by Statistica (Statsoft v. 5.0). The response surface methodology (RSM) had identified the optimum values where glutamine=2211.20 mg/L, glucose=1291.95 mg/L, and lipid mixtures 1000x=0.64 %v/v. Using the optimized parameters, the studies demonstrated an increase in the rhTf yield by three–fold from 19.89 μg/ml to 65.12 μg/ml.
Key words: Human transferrin; insect cells baculovirus; experimental design; central composite design; response surface methodology