Analysis of protein adsorption and binding at biosensor polymer interfaces using X-ray photon spectroscopy and scanning electrochemical microscopy.
We describe a method, based on X-ray photoelectron spectroscopy (XPS) measurements, to assess the extent of protein adsorption or binding on a variety of different muTAS and biosensor interfaces. Underpinning this method is the labeling of protein molecules with either iodine- or bromine-containing motifs by using protocols previously developed for radiotracer studies. Using this method, we have examined the adsorption and binding properties of a variety of modified electrodeposited polymer interfaces as well as other materials used in muTAS device fabrication. Using polymer interfaces modified with poly(propylene glycol) (PPG) chains, our results indicate that a chain of at least approximately 30 monomer units is required to inhibit nonspecific adsorption from concentrated protein solutions. The XPS methodology was also used to probe specific binding of avidins and enzyme conjugates thereof to biotinylated and mixed biotin/PPG-modified polymer interfaces. In one example, using competitive binding, it was established that the mode of binding of a peroxidase-streptavidin conjugate to a biotinylated modified polymer interface was primarily via the streptavidin moiety (as opposed to nonspecific binding via the enzyme conjugate). XPS evaluation of nonspecific and specific peroxidase-streptavidin immobilization on various functionalized polymers has guided the design and fabrication of functionalized interdigitated electrodes in a biosensing muTAS device. Subsequent characterization of this device using scanning electrochemical microscopy (SECM) corroborated the adsorption and binding previously inferred from XPS measurements on macroscale electrodes.
Glidle
Andrew
A
Bioelectronics Research Group, Department of Electronics and Electrical Engineering, University of Glasgow, Glasgow, G12 8LT, UK.
Yasukawa
Tomoyuki
T
Hadyoon
Charlotte S
CS
Anicet
Nathalie
N
Matsue
Tomokazu
T
Nomura
Masayuki
M
Cooper
Jon M
JM
D000327:Adsorption; D015374:Biosensing Techniques / Q000379:methods; D001710:Biotin / Q000737:chemistry; D004563:Electrochemistry / Q000295:instrumentation; D008853:Microscopy / Q000379:methods; D011108:Polymers / Q000737:chemistry*; D011409:Propylene Glycols / Q000737:chemistry*; D011506:Proteins / Q000737:chemistry*; D013052:Spectrometry, X-Ray Emission / Q000379:methods; D013499:Surface Properties
D011108:Polymers; D011409:Propylene Glycols; D011506:Proteins; C012504:polypropylene glycol; D001710:Biotin