Analysis of protein adsorption and binding at biosensor polymer interfaces using X-ray photon spectroscopy and scanning electrochemical microscopy

被引:34
作者
Glidle, A
Yasukawa, T
Hadyoon, CS
Anicet, N
Matsue, T
Nomura, M
Cooper, JM [1 ]
机构
[1] Univ Glasgow, Dept Elect & Elect Engn, Bioelect Res Grp, Glasgow G12 8LT, Lanark, Scotland
[2] Tohoku Univ, Grad Sch Engn, Dept Biomol Engn, Sendai, Miyagi 9808579, Japan
[3] Asahi Kasei Corp, Cent Technol Lab, Fuji, Shizuoka 4168501, Japan
关键词
D O I
10.1021/ac0261653
中图分类号
O65 [分析化学];
学科分类号
070302 ; 081704 ;
摘要
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 yTAS 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 pTAS device fabrication. Using polymer interfaces modified with poly(propylene glycol) (PPG) chains, our results indicate that a chain of at least similar to30 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.
引用
收藏
页码:2559 / 2570
页数:12
相关论文
共 35 条
[1]  
ALLEN RC, 1999, PROTEIN STAINING IDE
[2]   THE SYNTHESIS OF THYROXINE AND RELATED SUBSTANCES .7. THE PREPARATION OF DIPHENYL ETHERS FROM 2-6-DI-IODOPHENOLS [J].
BARNES, JH ;
BORROWS, ET ;
ELKS, J ;
HEMS, BA ;
LONG, AG .
JOURNAL OF THE CHEMICAL SOCIETY, 1950, (OCT) :2824-2833
[3]   LABELING OF PROTEINS TO HIGH SPECIFIC RADIOACTIVITIES BY CONJUGATION TO A I-125-CONTAINING ACYLATING AGENT - APPLICATION TO RADIOIMMUNOASSAY [J].
BOLTON, AE ;
HUNTER, WM .
BIOCHEMICAL JOURNAL, 1973, 133 (03) :529-538
[4]  
CHILKOTI A, 1993, SURFACE CHARACTERIZA
[5]   OXIDATIVE CLEAVAGE OF AMIDES - A METHOD FOR SELECTIVE CHEMICAL DEGRADATION OF PEPTIDES [J].
COREY, EJ ;
HAEFELE, LF .
JOURNAL OF THE AMERICAN CHEMICAL SOCIETY, 1959, 81 (09) :2225-2228
[6]  
Crowther J.R., 1995, ELISA: Theory and Practice, V42
[7]   Estimation of inelastic mean free paths for polymers and other organic materials: use of quantitative structure-property relationships [J].
Cumpson, PJ .
SURFACE AND INTERFACE ANALYSIS, 2001, 31 (01) :23-34
[8]   Protein adsorption on model surfaces with controlled nanotopography and chemistry [J].
Denis, FA ;
Hanarp, P ;
Sutherland, DS ;
Gold, J ;
Mustin, C ;
Rouxhet, PG ;
Dufrêne, YF .
LANGMUIR, 2002, 18 (03) :819-828
[9]   Formation and characterization of protein monolayers on oxygen-exposing surfaces by multiple-step self-chemisorption [J].
Facci, P ;
Alliata, D ;
Andolfi, L ;
Schnyder, B ;
Kötz, R .
SURFACE SCIENCE, 2002, 504 (1-3) :282-292
[10]  
FRAKER PJ, 1978, BIOCHEM BIOPH RES CO, V80, P849, DOI 10.1016/0006-291X(78)91322-0