Increased production of human granulocyte-macrophage colony stimulating factor (hGM-CSF) by the addition of stabilizing polymer in plant suspension cultures.
Granulocyte-macrophage colony-stimulating factor (GM-CSF) is a glycoprotein that stimulates the production of granulocytes, macrophages, and white blood cells. Secretion of human GM-CSF from cell suspension cultures of genetically modified tobacco has been facilitated using natural mammalian leader sequences. At the mid-exponential growth phase (day 4 after the initiation of cell suspension culture), GM-CSF was detected in the medium at a maximum concentration of 180 microg l(-1). However, the secreted GM-CSF was unstable in the medium, and rapidly degraded after day 5. In order to stabilize the secreted GM-CSF, three stabilizing polymers were tested, polyethylene glycol, polyvinylpyrrolidone and gelatin. Gelatin was the most effective in stabilizing the secreted GM-CSF. Following the addition of 5% (w/v) gelatin, the maximum GM-CSF concentration reached 783 microg l(-1), a 4.6-fold increase over control.
Lee
Jae-Hwa
JH
Basic Science Research Institute, Chonbuk National University, Dukjindong 664-14, Chonju, Chonbuk 561-756, South Korea.
Kim
Nan-Sun
NS
Kwon
Tae-Ho
TH
Jang
Yong-Suk
YS
Yang
Moon-Sik
MS
D016960:Agrobacterium tumefaciens / Q000737:chemistry / Q000235:genetics; D002455:Cell Division / Q000502:physiology; D018076:DNA, Complementary / Q000235:genetics; D010450:Endopeptidases / Q000032:analysis / Q000378:metabolism; D005079:Excipients / Q000378:metabolism; D005780:Gelatin / Q000378:metabolism*; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000096:biosynthesis* / Q000235:genetics / Q000378:metabolism; D006801:Humans; D011092:Polyethylene Glycols / Q000378:metabolism*; D011205:Povidone / Q000378:metabolism*; D014026:Nicotiana / Q000166:cytology* / Q000235:genetics / Q000378:metabolism* / Q000382:microbiology; D014169:Transformation, Bacterial
D018076:DNA, Complementary; D005079:Excipients; D011092:Polyethylene Glycols; D016178:Granulocyte-Macrophage Colony-Stimulating Factor; D005780:Gelatin; D010450:Endopeptidases; D011205:Povidone