Selection and Characterization of DNA Aptamers for Egg White Lysozyme

Molecules

Dinh T. Tran¹  Kris P. F. Janssen¹  Jeroen Pollet¹  Elke Lammertyn¹  Jozef Anné¹  Ann Van Schepdael¹ 

[1] ¹BIOSYST-MeBioS, Katholieke Universiteit Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium ²Laboratory of Bacteriology, Rega Institute, Katholieke Universiteit Leuven, Minderbroedersstraat 10, B-3000 Leuven, Belgium ³Laboratory for Pharmaceutical Analysis, O&N II Herestraat 49 - bus 923, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium

关键词: aptamer;    capillary electrophoresis;    food allergen;    lysozyme;    systematic evolution of ligands by exponential enrichment;   

DOI  :  10.3390/molecules15031127

来源: mdpi

【 摘 要 】

We have selected aptamers binding to lysozyme from a DNA library using capillary electrophoresis-systematic evolution of ligands by exponential enrichment. During the selection process the dissociation constant of the ssDNA pool decreased from the micromolar to the low nanomolar range within five rounds of selection. The final aptamer had a dissociation constant of 2.8 ± 0.3 nM, 6.1 ± 0.5 nM, and 52.9 ± 9.1 nM as determined by fluorescence anisotropy, surface plasmon resonance and affinity capillary electrophoresis respectively. The aptamers were successfully challenged for specificity against other egg white proteins. The high affinity aptamers open up possibilities for the development of aptamer based food and medical diagnostics.

【 授权许可】

CC BY   
This is an open access article distributed under the Creative Commons Attribution License (CC BY) which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.