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논문 기본 정보
- 자료유형
- 학술저널
- 저자정보
- 저널정보
- 한국미생물생명공학회 Journal of Microbiology and Biotechnology Journal of Microbiology and Biotechnology 제18권 제2호
- 발행연도
- 2008.1
- 수록면
- 383 - 391 (9page)
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The glycans linked to the insect cell-derived glycoproteins are known to differ from those expressed in mammalian cells, partly because of the low level or lack of glycosyltransferase activities. GnT II, GnT IV, GnT V, and ST3Gal IV, which play important roles in the synthesis of tetraantennarytype complex glycan structures in mammalian cells, were overexpressed in Trichoplusia ni cells by using a baculovirus expression vector. The glycosyltransferases, expressed as a fusion form with the IgG-binding domain, were secreted into the culture media and purified using IgG sepharose resin. The enzyme assay, performed using a pyridylaminatedsugar chain as an acceptor, indicated that the purified glycosyltransferases retained their enzyme activities. Human erythropoietin expressed in T. ni cells (rhEPO) was subjected to in vitro glycosylation by using recombinant glycosyltransferases and was converted into complex-type glycan with terminal sialic acid. The presence of Nacetylglucosamine, galactose, and sialic acid on the rhEPO moiety was detected by a lectin blot analysis, and the addition of galactose and sialic acid to rhEPO was confirmed by autoradiography using UDP-14C-Gal and CMP-14C-Sia as donors. The in vitro glycosylated rhEPO was injected into mice, and the number of reticulocytes among the red blood cells was counted using FACS. A significant increase in the number of reticulocytes was not observed in the mice injected with in vitro glycosylated rhEPO as compared with those injected with rhEPO.
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참고문헌 (22)
참고문헌 신청
Altmann, F. / 1993 / Processing of asparagines-linked oligosaccharides in insect cells. N-Acetylglucosaminyltransferase I and II activities in cultured lepidopteran cells / G
Altmann, F. / 1999 / Insect cells as host for the expression of recombinant glycoproteins / Glycoconj 16 : 109 ~ 123
Butters, T. D. / 1981 / Steps in the biosynthesis of mosquito cell membrane glycoproteins andthe effects of tunicamycin. Biochim. Biophys / Acta 640 : 672 ~ 686
Chang, K. H. / 2005 / Expression and characterization of recombinant β-secretase from Trichoplusia ni BTI Tn5B1-4 cells transformed with cDNAs encoding human β1,4- galactos
DEMIR, ISMAIL / 2006 / A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line / Journal of Microbiology and Biotechnology 16 (10) : 1
Altmann, F. / 1999 / Insect cells as host for the expression of recombinant glycoproteins / Glycoconj 16 : 109 ~ 123
Butters, T. D. / 1981 / Steps in the biosynthesis of mosquito cell membrane glycoproteins andthe effects of tunicamycin. Biochim. Biophys / Acta 640 : 672 ~ 686
Chang, K. H. / 2005 / Expression and characterization of recombinant β-secretase from Trichoplusia ni BTI Tn5B1-4 cells transformed with cDNAs encoding human β1,4- galactos
DEMIR, ISMAIL / 2006 / A Productive Replication of Hyphantria cunea Nucleopolyhedrovirus in Lymantria dispar Cell Line / Journal of Microbiology and Biotechnology 16 (10) : 1
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