Surface energy modified chips for detection of conformational states and enzymatic activity in biomolecules.
A novel patterning method for anchoring biomolecules and noncovalent assembled conjugated polyelectrolyte (CPE)/biomolecule complexes to a chip surface is presented. The surface energy of a hydrophilic substrate is modified using an elastomeric poly(dimethylsiloxane) (PDMS) stamp, containing a relief pattern. Modification takes place on the parts where the PDMS stamp is in conformal contact with the substrate and leaves low molecular weight PDMS residues on the surface resulting in a hydrophobic modification, and then biomolecules and CPE/biomolecule complexes are then adsorbed in a specific pattern. The method constitutes a discrimination system for different conformations in biomolecules using CPEs as reporters and the PDMS modified substrates as the discriminator. Detection of different conformations in two biomacromolecules, a synthetic peptide (JR2E) and a protein (calmodulin), reported by the CPE and resolved by fluorescence was demonstrated. Also, excellent enzyme activity in patterned CPE/horseradish peroxidase (HRP) enzyme was shown, demonstrating that this method can be used to pattern biomolecules with their activity retained. The method presented could be useful in various biochip applications, such as analyzing proteins and peptides in large-scale production, in making metabolic chips, and for making multi-microarrays.
Asberg
Peter
P
Biomolecular and Organic Electronics, IFM, Linköping University, S-581 83 Linköping, Sweden. petas@ifm.liu.se
Nilsson
K Peter R
KP
Inganäs
Olle
O
D002147:Calmodulin / Q000737:chemistry; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D004798:Enzymes / Q000737:chemistry*; D031541:Fluorescence Resonance Energy Transfer; D046912:Multiprotein Complexes; D009757:Nylons / Q000737:chemistry; D010455:Peptides / Q000737:chemistry; D040081:Protein Array Analysis; D011487:Protein Conformation; D013499:Surface Properties; D013816:Thermodynamics
D002147:Calmodulin; D004129:Dimethylpolysiloxanes; D004798:Enzymes; D046912:Multiprotein Complexes; D009757:Nylons; D010455:Peptides; C085707:poly(dimethylsiloxane)-polyamide copolymer