Recombinant human granulocyte macrophage colony stimulating factor (rhGM -CSF), a glycoprotein induces proliferation of myeloid progenitor cells (1, 2), is produced from a variety of organisms such as Chinese hamster ovary, Saccharomyces cervisiae, E. coli, and Oryza sativa. rrhGM-CSF (rice cell origin, Oryza sativa) was recently shown to be more stable in-vivo than rhGM-CSF produced from different origins (3). However, in-vitro stability of rrhGM-CSF in liquid state has not been optimized, so the activity of rrhGM-CSF incubated at room temperature decreased more than 85% after 8 weeks. Heat treatment on rrhGM-CSF was carried out to increase stability, and rrhGM-CSF activity was evaluated by measuring proliferation rate of TF-1 cells (human pre-myeloid cell line). The activity of heat-treated rrhGM-CSF for 1, 2, 4, 6 min at 70℃ was preserved as 15%, 50%, 78%, 100% (± 20%) after 8 weeks, respectively. Protease inhibitor cocktail (Roche) also increased the stability of rrhGM-CSF. Therefore, reduction of rrhGM-CSF activity in liquid state is estimated due to plant-derived protease. The stability of heat-treated rrhGM-CSF was also affected by the degree of glycosylation. In the thermal stability of heat-treated rrhGM-CSF at 100℃, 30 min, high-glycosylated rrhGM -CSF was higher than low-glycosylated rrhGM-CSF. However, the stability was not different between high-glycosylated and low-glycosylated rrhGM-CSF in the case of heat treatment at 100℃ above 30 min. In this study, we increased the stability of rrhGM-CSF in liquid state by heat treatment, and confirmed that the thermal stability was affected by the degree of glycosylation of rrhGM-CSF.
키워드
GM-CSFproteaseGM-CSF stabilityglycosylation
저자
Chang-Koo YUN [ Dept. of Applied Bioscience, CHA University, Seongnam-si, Gyeonggi-do 463-840, Korea. ]
Kyu-Boem HAN [ CHABIO&DIOSTEH Co., Ltd., 533 (1-103) Yongsan-dong, Yuseong-gu, Daejeon 305-500, Korea. ]
Youn-Je PARK [ Dept. of Applied Bioscience, CHA University, Seongnam-si, Gyeonggi-do 463-840, Korea. ]
한국생물공학회 [The Korean Society for Biotechnology and Bioengineering]
설립연도
1984
분야
공학>생물공학
소개
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