Extraction and purification of human interleukin-10 from transgenic rice seeds.
Recombinant protein production system using transgenic rice grain offers many advantages in higher accumulation, preservation, lower production cost, ease of scale up and low risk of contamination by toxic materials. We developed a transgenic rice strain whose seeds accumulate human interleukin (IL)-10, a cytokine that suppresses inflammation-related immune responses. We also developed a method of extracting and purifying IL-10 from rice seeds. A biochemical crosslinking method was used to detect the biologically active noncovalent dimer of IL-10. This method was useful for developing efficient methods of refolding and purification. The purified IL-10 comprised only noncovalent dimers and showed higher activity than the commercial IL-10. The purified IL-10 had very low endotoxin contamination and is expected to have broad clinical application.
Preventec, Inc., 1-2 Owashi, Tsukuba, Ibaraki 305-8634, Japan. fwara@preventec-inc.com
Yoshihiro
Y
Fujiwara
Yasuhiko
Y
Aiki
Lijun
L
Yang
Fumio
F
Takaiwa
Akemi
A
Kosaka
Noriko M
NM
Tsuji
Kentaro
K
Shiraki
Kenji
K
Sekikawa
D000818:Animals; D002478:Cells, Cultured; D018076:DNA, Complementary / Q000235:genetics; D003713:Dendritic Cells / Q000276:immunology; D006801:Humans; D016753:Interleukin-10 / Q000737:chemistry / Q000235:genetics* / Q000276:immunology / Q000302:isolation & purification*; D051379:Mice; D008807:Mice, Inbred BALB C; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D017510:Protein Folding; D055503:Protein Multimerization; D012639:Seeds / Q000235:genetics*
D018076:DNA, Complementary; D016753:Interleukin-10