KIT | KIT-Bibliothek | Impressum

High affinity immobilization of proteins using the CrAsH/TC tag

Schulte-Zweckel, J.; Rosi, F.; Sreenu, D.; Schröder, H.; Niemeyer, C.M.; Triola, G.



Abstract (englisch): Protein microarrays represent important tools for biomedical analysis. We have recently described the use of the biarsenical-tetracysteine (TC) tag for the preparation of protein microarrays. The unique feature of this tag enables the site-specific immobilization of TC-containing proteins on biarsenical-modified surfaces, resulting in a fluorescence enhancement that allows the direct quantification of the immobilized proteins. Moreover, the reversibility of the binding upon incubation with large quantities of thiols permits the detachment of the proteins from the surface, thereby enabling recovery of the substrate to extend the life time of the slide. Herein, we describe our recent results that further extend the applicability of the CrAsH/TC tag to the fabrication of biochips. With this aim, the immobilization of proteins on surfaces has been investigated using two different spacers and two TC tags, the minimal TC sequence (CCPGCC) and an optimized motif (FLNCCPGCCMEP). While the minimal peptide motif enables a rapid recycling of the slide, the optimized TC sequence reveals an increased affinity due to its greater resistance to displacement by thiols. Moreover, the developed methodology was applied to the immobilization of proteins via on-chip ligation of recombinant protein thioesters.


Zugehörige Institution(en) am KIT Institut für Biologische Grenzflächen (IBG)
Publikationstyp Zeitschriftenaufsatz
Jahr 2016
Sprache Englisch
Identifikator DOI: 10.3390/molecules21060750
ISSN: 1420-3049
URN: urn:nbn:de:swb:90-577459
KITopen ID: 1000057745
HGF-Programm 47.02.01; LK 01
Erschienen in Molecules
Band 21
Heft 6
Seiten 750
Schlagworte protein microarray, protein immobilization, FlAsH, CrAsH, site-selective, on-chip ligation
KIT – Die Forschungsuniversität in der Helmholtz-Gemeinschaft KITopen Landing Page