Denaturing for Nanoarchitectonics: Local and Periodic UV-Laser Photodeactivation of Protein Biolayers to Create Functional Patterns for Biosensing

Abstract

The nanostructuration of biolayers has become a paradigm for exploiting nanoscopic light-matter phenomena for biosensing, among other biomedical purposes. In this work, we present a photopatterning method to create periodic structures of biomacromolecules, based on a local and periodic mild denaturation of protein biolayers mediated by UV-laser irradiation. These nanostructures are constituted by a periodic modulation of the protein activity, so they are free of topographic and compositional changes along the pattern. Herein we introduce the approach, explore the patterning parameters, characterize the resulting structures, and assess their overall homogeneity. This UV-based patterning principle has proven to be an easy, cost-effective, and fast way to fabricate large areas of homogeneous one-dimensional protein patterns (2 min, 15 x 1.2 mm, relative standard deviation ≃ 16%). This work also investigates the implementation of these protein patterns as transducers for 2 diffractive biosensing. Using a model immunoassay, these patterns have demonstrated negligible signal contributions from non-specific bindings and comparable experimental limits of detection in buffer medium and in human serum (53 and 36 ng•mL -1 of unlabelled IgG, respectively).

Microstructured molecular BIO-gratings by means of UV induced denaturation - INVITED

“…We have presented a reliable, rapid fabrication method of BIO-gratings for biosensing, with enhanced performance compared to other techniques, for example microcontact printing, as it is detailed in [7].…”

“…Complete immunoassays were performed for the optimized BIO-gratings in PBS buffer dilutions of different concentrations of aBSA, as well as in human serum dilutions. The DL and quantification limits of the biosensors are 36 ng/mL and 100 ng/mL (aBSA in human serum), and 53 ng/mL and 164 ng/mL (aBSA in PBS buffer) [7]. It is also worth to note that the BIO-gratings remained active for at least 30 days stored at 4ºC.…”

Resonant Couplings in U-Shaped Fibers for Biosensing

“…As said before, the calculation of sensitivity and detection limit presented here arises from a conservative assumption. From previous works [10,30] we know that the linear range of BSA/aBSA biosensor ends around 10 g•mL -1 , but since those transducers rely on different operation principle (diffractive grating), we chose a aBSA concentration that was further indeed of that limit, to be sure that we were observing the maximum possible response of the biosensor. Thus we can expect that a more detailed study would provide detection limits at least a factor of 5 better than this estimate.…”

Materials Nanoarchitectonics for Advanced Physics Research

“…The concept of nanoarchitectonics can be applied to various fields from basic to applied. The nanoarchitectonics approaches are used in chemistry such as material synthesis, [39,40] structure control, [41,42] and catalysis, [43,44] in biology such as basic biochemistry [45,46] and biomedical sciences, [47,48] in functional instrumental systems such as sensors [49,50] and devices, [51,52] and in practical applications such as the environment applications [53,54] and energy applications. [55,56] Although many of these may appear to be chemistry or biology in nature, the underlying phenomena are based on physical principles.…”