Label-Free Measurement of Amyloid Elongation by Suspended Microchannel ResonatorsClick to copy article linkArticle link copied!
Abstract
Protein aggregation is a widely studied phenomenon that is associated with many human diseases and with the degradation of biotechnological products. Here, we establish a new label-free method for characterizing the aggregation kinetics of proteins into amyloid fibrils by suspended microchannel resonators (SMR). SMR devices are unique in their ability to provide mass-based measurements under reaction-limited conditions in a 10 pL volume. To demonstrate the method, insulin seed fibrils of defined length, characterized by atomic force microscopy (AFM) and transmission electron microscopy (TEM), were covalently immobilized inside microchannels embedded within a micromechanical resonator, and the elongation of these fibrils under a continuous flow of monomer solution (rate ∼1 nL/s) was measured by monitoring the resonance frequency shift. The kinetics for concentrations below ∼0.6 mg/mL fits well with an irreversible bimolecular binding model with the rate constant kon = (1.2 ± 0.1) × 103 M–1 s–1. Rate saturation occurred at higher concentrations. The nonlinear on-rate for monomer concentrations from 0 to 6 mg/mL and for temperatures from 20 to 42 °C fit well globally with an energy landscape model characterized by a single activation barrier. Finally, elongation rates were studied under different solution conditions and in the presence of a small molecule inhibitor of amyloid growth. Due to the low volume requirements, high precision, and speed of SMR measurements, the method may become a valuable new tool in the screening for inhibitors and the study of fundamental biophysical mechanisms of protein aggregation processes.
Cited By
This article is cited by 12 publications.
- Wen Wang, Jiaxing Zhang, Wei Qi, Rongxin Su, Zhimin He, Xin Peng. Alizarin and Purpurin from Rubia tinctorum L. Suppress Insulin Fibrillation and Reduce the Amyloid-Induced Cytotoxicity. ACS Chemical Neuroscience 2021, 12
(12)
, 2182-2193. https://doi.org/10.1021/acschemneuro.1c00177
- Rafita Erli Adhawiyah, Jungchul Lee. Enhancing Sensitivity and Selectivity: Morphological Modification and Chemical Functionalization within Confined Structures. International Journal of Precision Engineering and Manufacturing 2024, 25
(4)
, 875-895. https://doi.org/10.1007/s12541-023-00935-z
- Tao Ma, Anle Mu. Study on the Stability of Functionally Graded Simply Supported Fluid-Conveying Microtube under Multi-Physical Fields. Micromachines 2022, 13
(6)
, 895. https://doi.org/10.3390/mi13060895
- Juhee Ko, Jaewoo Jeong, Sukbom Son, Jungchul Lee. Cellular and biomolecular detection based on suspended microchannel resonators. Biomedical Engineering Letters 2021, 11
(4)
, 367-382. https://doi.org/10.1007/s13534-021-00207-7
- Julia Gross-Rother, Michaela Blech, Eduard Preis, Udo Bakowsky, Patrick Garidel. Particle Detection and Characterization for Biopharmaceutical Applications: Current Principles of Established and Alternative Techniques. Pharmaceutics 2020, 12
(11)
, 1112. https://doi.org/10.3390/pharmaceutics12111112
- Davide Scaiola, Stefano Stassi, Roberta Calmo, Damien Maillard, Stefano S.G. Varricchio, Annalisa De Pastina, Guillermo Villanueva, Philippe Renaud, Carlo Ricciardi. Fabrication of clamped-clamped beam resonators with embedded fluidic nanochannel. Microelectronic Engineering 2020, 231 , 111395. https://doi.org/10.1016/j.mee.2020.111395
- Andrea Hawe, Daniel Weinbuch, Sarah Zölls, Angelika Reichel, John F. Carpenter. Submicrometer, micrometer and visible particle analysis in biopharmaceutical research and development. 2020, 285-310. https://doi.org/10.1016/B978-0-444-64173-1.00010-X
- Roberta Calmo, Andrea Lovera, Stefano Stassi, Alessandro Chiadò, Davide Scaiola, Francesca Bosco, Carlo Ricciardi. Monolithic glass suspended microchannel resonators for enhanced mass sensing of liquids. Sensors and Actuators B: Chemical 2019, 283 , 298-303. https://doi.org/10.1016/j.snb.2018.12.019
- Manizhe Zakeri, Seyed Mahmoud Seyedi Sahebari. Modeling and simulation of a suspended microchannel resonator nano-sensor. Microsystem Technologies 2018, 24
(2)
, 1153-1166. https://doi.org/10.1007/s00542-017-3478-6
- Han Yan, Wen-Ming Zhang, Hui-Ming Jiang, Kai-Ming Hu. Pull-In Effect of Suspended Microchannel Resonator Sensor Subjected to Electrostatic Actuation. Sensors 2017, 17
(12)
, 114. https://doi.org/10.3390/s17010114
- Han Yan, Wen-Ming Zhang, Hui-Ming Jiang, Kai-Ming Hu, Fang-Jun Hong, Zhi-Ke Peng, Guang Meng. A measurement criterion for accurate mass detection using vibrating suspended microchannel resonators. Journal of Sound and Vibration 2017, 403 , 1-20. https://doi.org/10.1016/j.jsv.2017.05.030
- Wen-Ming Zhang, Han Yan, Hui-Ming Jiang, Kai-Ming Hu, Zhi-Ke Peng, Guang Meng. Dynamics of suspended microchannel resonators conveying opposite internal fluid flow: Stability, frequency shift and energy dissipation. Journal of Sound and Vibration 2016, 368 , 103-120. https://doi.org/10.1016/j.jsv.2016.01.029
Article Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.
Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.
The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated.