Ribosome Display

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Ribosome Display in Antibody Selection
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Ribosome Display ??? ???? ?????? ??????? ???? 6
???? ?????? ????? RNA-protein fusion. ????? ???
??????? ?????? ?????? ?? ??????? ???? ?????
????? ???????? ?????. ????? ??? ????????
???? ??????-????? (VEGF ???????? ?? ???
???????) ??????-????? (?? ????? ?????? ????
????????)
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????? ??? ????? ??????? ?? ???? ??? 1. ?? ?????
??????? ???????? ??????? ???? ?????? ??? in
vivo- ????? ?? ???? ?????? ????? ??????????
?? ?????????.
?? ?????? in vitro
2. ?????? ????? ?? ???? ????? ???? ?????
????????? ?? ??? ????? ?? ?- clone ????? ??
????? ????? ?????? ????? ?????.
???? ????? ??? ??????? ???????.
3. Protein Evolution-??????? ?????? ????? ?????
?????? ??????- Affinity Maturation
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???? ???? ?????? ???????
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??? ?????????
SsrA
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?? ??? ????? ???? Ribosome Display ?? ????? ??
1. ???? ??- mRNA ????? ?? ????????
?. ????? ?? ????? stop/ ?. ????? ?- RNA ?? ????
stemloop ?. ????? ?????? RNAse
2. ???? ?????? ????? ?? ???????? ?. ????? ??
????? stop ?. ???? ?? SsrA 3. ??????
???? ?????? ????? ??? ??? ?????? ???????? ?.
???? ??? ?? ?????? ?????? ?. ????? ????????
?. C-terminal linker
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?? ??? ?????? ????? ???? ?? ?????? ???? ???? ????
??????? ??????? ?? ?- DNA ????? ??
T7
SD
VL
VH
spacer
tag
linker
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???? ????? ???? Ribosome Display?
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????? ????? ?????? ????? ?? ???? ?????? In
vitro selection and evolution of functional
proteins by using ribosome display Jozef Hanes
Andreas Pluckthun 1997
????? ??? scFv ?? ????? ???? hemagglutinin. ?????
???? ?? ????? ?- stemloop ???? ????? ?-mRNA. (???
?? ?- RNA ???? ????? ?? RNAse)
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???? ?- mRNA ?? scFvhag ?????? ????????
????? ????? ??????? ?? ??????? ?????? ???????
????? ??? ?????? ?????? ????????.
?? ???? ?- stemloop ???? ??? ?- RNA ???????? ????
????? ????? ????? ????????? ?? RNA.
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????? ?? ???????? scFvhag ribosome ???????
?? scFval2
????? ???????? scFvhag scFval2 1108
???? ????? ?????? ?????? ??????? hag ?90
????????? ?????????? ????? scFvhag . ?????? ???
????? ????? ?? 2 ???? ???? ?? ????? ??????.
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????? ???????? ????? ???????
tr- ???????? ??????? amp- ?????????-???????? hag-
??????????-????????
scFvhag ?????? ???? ????? ?? ???? ?????? ??????
?? hag ???????.
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20 clones ?????
2 ????? scFval2
18 ????? scFvhag
13 ???? 43-102 ???????
5 ???? lt18 ???????
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?????? 1. ???? ?????? ????? Ribosome Display
???? ????? ??????? ????? ??????? ????. 2.
????? ?? ?????? ??????? ??????? ?????? ??
???????? ??? ???????? ?? ????? ???????? error
prone ????? ??????? ????????? ??? ?????. 3.?????
???? ??? ?? ???????????, ????? ????? ????-
??? ????? ???????? ?? ??????? ??????. 4. ???????
????????? ?????? ???? - ?? 1015 ???????
????? ????? ??? ???? ????.