Nonspecific binding removal from protein microarrays using thickness shear mode resonators

Abstract

Nonspecific binding is a universal problem that reduces bioassay sensitivity and specificity. We demonstrate that ultrasonic waves, generated by 5-MHz quartz crystal resonators, accelerate nonspecifically bound protein desorption from sensing and nonsensing areas of micropatterned protein arrays, controllably and nondestructively cleaning the micropatterns. Nonsensing area fluorescent intensity values dropped by more than 85% and sensing area fluorescent intensity dropped 77% due to nonspecific binding removal at an input power of 14 W. After patterning, antibody films were many layers thick with nonspecifically bound protein, and aggregates obscured patterns. Quartz crystal resonators removed excess antibody layers and aggregates leaving highly uniform films, as evidenced by smaller spatial variations in fluorescent intensity and atomic force microscope surface roughness values. Fluorescent intensity values obtained after 14-W QCR operation were more repeatable and uniform.Index Terms-Nonspecific binding, protein microarray, quartz crystal resonator, ultrasonic.

Chitosan cushioned phospholipid membrane and its application in imaging ellipsometry based-biosensor

“…Proteins immobilized directly on silicon surface have been successfully used to capture biomarkers [3,4], phage [5] or virus [6,7]. Aiming at protein detection in complex biological samples, much attention should be paid to avoid non-specific protein adsorption, which is the main source of noise in immunoassays and has become the major obstacle in developing more sensitive and higher throughput biosensor assays [8][9][10][11].…”

An investigation of vibration-induced protein desorption mechanism using a micromachined membrane and PZT plate

“…Examples include polymer-based coatings (Gref et al 2000;Huang et al 2001;Nolan et al 2005;Nuzzo 2003; Prime and Whitesides 1993;Allen et al 2002;Doherty et al 2003;Jacobs et al 1988;Makamba et al 2003;Whelan and Regan 2006), surface functionalization using gas plasma (Sigal et al 1998), surface charges (Pasche et al 2005), detergent coating (Zhang and Ferrari 1997), nano-structured surfaces (Ainslie et al 2005;Rigby et al 1995;Rigby et al 1996), and mechanical vibration (Ainslie et al 2005;Meyer et al 2006;Rahmoune and Latour 1995;Yeh et al 2007). Among these methods, passive methods (without surface treatment) are more attractive for certain applications (long-circulating sterically protected liposome, Klibanov et al 2003;biosensor, Sharma et al 2004;protein chip, Zhu and Snyder 2003).…”

“…The use of mechanical means to reduce non-specific protein binding has been investigated by researchers (Ainslie et al 2005;Meyer et al 2006;Rahmoune and Latour 1995). It was observed that magnetic nanowires, when oscillated by an external oscillating magnetic field, adsorb less proteins than static nanowires (Ainslie et al 2005).…”

“…It was observed that magnetic nanowires, when oscillated by an external oscillating magnetic field, adsorb less proteins than static nanowires (Ainslie et al 2005). Using piezoelectric materials in the construction of the sensor is another approach to combat biofouling (Meyer et al 2006;Rahmoune and Latour 1995). On an oceanographic sensor, the application of a mechanical wave induced by polyvinylidene fluoride (PVDF) was shown to reduce biofouling (Rahmoune and Latour 1995).…”

“…On an oceanographic sensor, the application of a mechanical wave induced by polyvinylidene fluoride (PVDF) was shown to reduce biofouling (Rahmoune and Latour 1995). High shear waves from high-frequency (~MHz) macro-scale quartz resonators (thickness-shear mode) with a 3.5 W output can attenuate non-specific protein binding (Meyer et al 2006). An external fluid flowing through a surface has also been shown to attenuate the adsorption of surface colloids and biomolecules (Qi and Brereton 1995;Rigby et al 1996).…”