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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
초록

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).

MATERIALS
제품 번호
브랜드
제품 설명

Sigma-Aldrich
Heparin sodium salt from porcine intestinal mucosa, ≥180 USP units/mg
Sigma-Aldrich
L-Methionine, reagent grade, ≥98% (HPLC)
Sigma-Aldrich
Adenosine 3′,5′-cyclic monophosphate sodium salt monohydrate, ≥98.0% (HPLC), powder
Sigma-Aldrich
Guanosine 5′-triphosphate sodium salt hydrate, ≥95% (HPLC), powder
Sigma-Aldrich
Dulbecco′s Modified Eagle′s Medium/Nutrient Mixture F-12 Ham, With L-glutamine, 15 mM HEPES, and sodium bicarbonate, liquid, sterile-filtered, suitable for cell culture
Sigma-Aldrich
Adenosine 5′-triphosphate disodium salt hydrate, Grade I, ≥99%, from microbial
Sigma-Aldrich
Uridine 5′-triphosphate trisodium salt hydrate, from yeast, Type III, ≥96%
Sigma-Aldrich
Lithium potassium acetyl phosphate, high-energy phosphate donor
Sigma-Aldrich
Anti-Mouse IgG (whole molecule)−Alkaline Phosphatase antibody produced in goat, affinity isolated antibody, buffered aqueous glycerol solution