Selection and Characterization of DNA Aptamers for Egg White Lysozyme 英文参考文献.docVIP

Molecules 2010, 15, 1127-1140; doi:10.3390/moleculeOPEN ACCESS molecules ISSN 1420-3049 /journal/molecules Article Selection and Characterization of DNA Aptamers for Egg White Lysozyme Dinh T. Tran 1, Kris P. F. Janssen 1, Jeroen Pollet 1, Elke Lammertyn 2, Jozef Anné 2, Ann Van Schepdael 3 and Jeroen Lammertyn 1,* 1 BIOSYST-MeBioS, Katholieke Universiteit Leuven, Willem de Croylaan 42, B-3001 Leuven, Belgium; E-Mails: ThiDinh.Tran@biw.kuleuven.be (D.T.T.); Kris.Janssen@biw.kuleuven.be (K.P.F.J.); Jeroen.Pollet@biw.kuleuven.be (J.P.) 2 Laboratory of Bacteriology, Rega Institute, Katholieke Universiteit Leuven, Minderbroedersstraat 10, B-3000 Leuven, Belgium; E-Mails: Elke.Lammertyn@lrd.kuleuven.be (E.L.); Jozef.Anne@rega.kuleuven.be (J.A.) 3 Laboratory for Pharmaceutical Analysis, ON II Herestraat 49 - bus 923, Katholieke Universiteit Leuven, B-3000 Leuven, Belgium; E-Mail: Ann.VanSchepdael@pharm.kuleuven.be (A.V.S.) ? Author to whom correspondence should be addressed; E-Mail: jeroen.lammertyn@biw.kuleuven.be; Tel.: + 32-16-321459; Fax: + 32-16-321614. Received: 13 January 2010; in revised form: 10 February 2010 / Accepted: 1 March 2010 / Published: 2 March 2010 Abstract: 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 di