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  • Ribosome display: a technology for selecting and evolving proteins from large libraries.

Ribosome display: a technology for selecting and evolving proteins from large libraries.

Methods in molecular biology (Clifton, N.J.) (2010-10-23)
Birgit Dreier, Andreas Plückthun
要旨

The selection and concomitant affinity maturation of proteins to bind to user-defined target molecules have become a key technology in biochemical research, diagnostics, and therapy. One of the most potent selection technologies for such applications is ribosome display. It works entirely in vitro, and this has two important consequences. First, since no transformation of any cells is required, libraries with much greater diversity can be handled than with most other techniques. Second, since a library does not have to be cloned and transformed, it is very convenient to introduce random errors in the library by PCR-based methods and select improved binders. Thus, a true directed evolution, an iteration between randomization and selection over several generations, can be conveniently carried out, e.g., for affinity maturation. Ribosome display has been used successfully for the selection of antibody fragments and other binding proteins, such as Designed Ankyrin Repeat Proteins (DARPins).

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Sigma-Aldrich
ダルベッコ改変イーグル培地/栄養混合物F-12ハム, With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
グアノシン 5′-三リン酸 ナトリウム塩 水和物, ≥95% (HPLC), powder
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アデノシン 5'-三リン酸 二ナトリウム塩 水和物, Grade I, ≥99%, from microbial
Sigma-Aldrich
(−)-アデノシン3′5′-サイクリック一リン酸 ナトリウム塩 一水和物, ≥98.0% (HPLC), powder
Sigma-Aldrich
ウリジン 5′-三リン酸 三ナトリウム塩 水和物, from yeast, Type III, ≥96%
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抗マウスIgG (全分子)−アルカリフォスファターゼ ヤギ宿主抗体, affinity isolated antibody, buffered aqueous glycerol solution
Sigma-Aldrich
アセチルリン酸リチウムカリウム, high-energy phosphate donor
Sigma-Aldrich
L-メチオニン, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
ヘパリン ナトリウム塩 from porcine intestinal mucosa, ≥180 USP units/mg