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An universal biolinker for immobilization of protein and oligoDNA on a glass slide chip

An universal biolinker for immobilization of protein and oligoDNA on a glass slide chip Abstract We have previously shown that a Calixarene derivative was an excellent agent for protein immobilization on a solid phase. We further characterized the recognition mechanism between the ‘host’ Calixarene derivative and ‘guest’ molecules in this study. The rate of complex formation of Calixcrown with amino acids informed that Calixcrown had the strongest interaction with Arg among the tested amino acids. In order to confirm that Arg is an effective residue in immobilization of protein on Calixcrown-coated glass, several human recombinant superoxide dismutase tagged with different numbers of Arg or Lys were generated. The recombinant protein tagged with longer Arg was immobilized better than the ones tagged with shorter Arg or Lys. We also tested if Calixcrown can immobilize oligoDNA as efficiently as it does protein. Calixcrown most strongly captured homo-dGTP oligoDNA among the four homo-oligoDNAs. When Calixcrown chip was applied to detect single nucleotide polymorphism, it differentiated one base pair mismatch of the double helix oligoDNA. Thus, Calixarene derivative linker molecule can be used as an efficient immobilization agent for both protein and oligoDNA with the right orientation on a solid phase. http://www.deepdyve.com/assets/images/DeepDyve-Logo-lg.png BioChip Journal Springer Journals

An universal biolinker for immobilization of protein and oligoDNA on a glass slide chip

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References (23)

Publisher
Springer Journals
Copyright
2012 The Korean BioChip Society and Springer-Verlag Berlin Heidelberg
ISSN
1976-0280
eISSN
2092-7843
DOI
10.1007/s13206-012-6101-z
Publisher site
See Article on Publisher Site

Abstract

Abstract We have previously shown that a Calixarene derivative was an excellent agent for protein immobilization on a solid phase. We further characterized the recognition mechanism between the ‘host’ Calixarene derivative and ‘guest’ molecules in this study. The rate of complex formation of Calixcrown with amino acids informed that Calixcrown had the strongest interaction with Arg among the tested amino acids. In order to confirm that Arg is an effective residue in immobilization of protein on Calixcrown-coated glass, several human recombinant superoxide dismutase tagged with different numbers of Arg or Lys were generated. The recombinant protein tagged with longer Arg was immobilized better than the ones tagged with shorter Arg or Lys. We also tested if Calixcrown can immobilize oligoDNA as efficiently as it does protein. Calixcrown most strongly captured homo-dGTP oligoDNA among the four homo-oligoDNAs. When Calixcrown chip was applied to detect single nucleotide polymorphism, it differentiated one base pair mismatch of the double helix oligoDNA. Thus, Calixarene derivative linker molecule can be used as an efficient immobilization agent for both protein and oligoDNA with the right orientation on a solid phase.

Journal

BioChip JournalSpringer Journals

Published: Mar 1, 2012

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