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

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.