AIR™ Technology

AIR™ is a revolutionary optical, label-free technology that enables biomarker discovery and research on the ZIVA Platform.
AIR™ stands for Arrayed Imaging Reflectometry and allows for increased plex and sensitivity. Adarza has been able to demonstrate the use of AIR™ to detect a variety of biological targets including small proteins, antibodies, nucleic acids, bacteria, and even viruses.  In addition, we have been able to utilize AIR™ to examine proteins or antibodies expressed at high endogenous levels and low levels.
How AIR™ Technology Works
AIR™ is a newly developed label-free optical biosensing technique based on the creation and perturbation of a condition of zero reflectance on a silicon substrate.
The antireflective coating is formed by covalently immobilizing arrayed probes (i.e. antibodies) on a silicon dioxide film. With no probe-target binding events, zero reflectance is generated when exposed to a laser beam. When probe-target complex forms (i.e. antibody-antigen), this results in a localized increase in optical thickness due to the increase in mass and a measurable reflectance change that is concentration/mass dependent. Using miniature silicon biosensors (approximately 5 x 5 mm), hundreds of probe spots can be arrayed and up to 100 targets can be interrogated. The biosensors can further be assembled in a 96-well format to enable high throughput of samples and automation (Figure 1).

Figure 1. Basic principle of AIR™ technology to achieve large density multiplex arrays. A) Using a silicon dioxide substrate with thickness specific to an array of antibody probes, near-zero reflectance is achieved when illuminated with s-polarized light at a fixed wavelength and angle B) Upon antigen bind­ing to unique antibodies arrayed on individual spots, concentration dependent signal is detected using a CCD camera. Further enhancement of signal and increased sensitivity can be achieved by adding secondary antibodies or any large target enhancers that result in increased mass C) Individual biosensors can be assembled into a 96-well format that can be further be automated in a detection system (ZIVA) for complete automation.

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References
  1. Mace, C. R., Striemer, C. C., Miller, B. L., “A theoretical and experimental analysis of Arrayed Imaging Reflectometry as a sensitive proteomics technique” Anal. Chem., 78, 5578-5583 (2006).
  2. Carter, J. A., Mehta, S. D., Mungillo M.V., Striemer, C. C., Miller, B. L., “Analysis of inflammatory biomarkers by Arrayed Imaging Reflectometry” Biosensors and Bioelectronics, 26, 3944-3948 (2011).
  3. Bucukovski, J.; Carter, J. A.; Striemer, C. C.; Müllner, S.; Schulte-Pelkum, J.; Schulz-Knappe, P.; Miller, B. L. “Label-free microarray-based detection of autoantibodies in human serum” J. Immunol. Methods, 459, 44-49 (2018).
  4. Zhang, H.; Henry, C.; Anderson, C. S.; Nogales, A.; DeDiego, M. L.; Bucukovski, J.; Martinez-Sobrido, L.; Wilson, P. C.; Topham, D. J.; Miller, B. L. “Crowd on a chip: Label-free human monoclonal antibody arrays for serotyping influenza”, Analytical Chemistry, 90, 9583-9590 (2018)
  5. Bucukovski, J.; Latorre-Margalef, N.; Stallknecht, D. E.; Miller, B. L. “A multiplex label-free approach to avian influenza surveillance and serology”, PLOS ONE, 10, e0134484 (2015)