New Generation Affibody Molecule Phage Library for Highly Stringent Binder Selections
Lin, Dong (2014)
2014
Master's Degree Programme in Science and Bioengineering
Luonnontieteiden tiedekunta - Faculty of Natural Sciences
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Hyväksymispäivämäärä
2014-06-04
Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi:tty-201405231206
https://urn.fi/URN:NBN:fi:tty-201405231206
Tiivistelmä
Affibody is a new class of engineered affinity protein, and has great properties com
pared with antibody. The aim of the project is to construct a new generation Affibody molecule phage library in which the bound phages are cleaved by trypsin in the selections against target molecules. First, a new phagemid vector pAffi-100-Tryp was created for new library preparation. Second, in comparative test selections with low-pH acid and trypsin cleavage elution, the functional display was ensured. Then, a new generation Affibody molecule library was constructed for highly stringent binder selections. The size of the library is approximately 4.7×10^9 cfu. In the selection against rhEphrin-B3, the bound phages were able to be cleaved by trypsin, and re-infect. After four rounds selection, candidate binders were selected for subcloning. According to the sequencing results, there were similar patterns among those candidate binders. Also, only Ile was found in Position 31.
pared with antibody. The aim of the project is to construct a new generation Affibody molecule phage library in which the bound phages are cleaved by trypsin in the selections against target molecules. First, a new phagemid vector pAffi-100-Tryp was created for new library preparation. Second, in comparative test selections with low-pH acid and trypsin cleavage elution, the functional display was ensured. Then, a new generation Affibody molecule library was constructed for highly stringent binder selections. The size of the library is approximately 4.7×10^9 cfu. In the selection against rhEphrin-B3, the bound phages were able to be cleaved by trypsin, and re-infect. After four rounds selection, candidate binders were selected for subcloning. According to the sequencing results, there were similar patterns among those candidate binders. Also, only Ile was found in Position 31.