Review
Chemistry, Analytical
Filip Dusa, Anna Kubesova, Jiri Salplachta, Dana Moravcova
Summary: Capillary isoelectric focusing (cIEF) is a powerful separation technique that requires the repeatable formation of a pH gradient and the use of internal standards to track its profile. This review provides an overview of available isoelectric point markers and their syntheses, as well as recent applications of cIEF in analyzing complex larger structures.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2023)
Article
Chemistry, Analytical
Junjie Zhang, Xuemeng Tian, Xihui Cui, Aqun Zheng, Jianjun Li, Yanhong Bai, Yuansuo Zheng
Summary: The separation and purification of proteins is crucial for proteomics research. This study presents a facile method to synthesize magnetic nanomaterials with dendritic structure and unique electrostatic effect, enabling selective adsorption of proteins. The materials exhibit stable crystal shape, superparamagnetic properties, fast adsorption kinetics, high adsorption capacity, and reusability, showing great potential for special protein adsorption.
Article
Chemistry, Multidisciplinary
Xiao Fan, Xiaoyu Zhang, Jinglei Ping
Summary: This article discusses a method for achieving transverse isoelectric focusing and electrokinetic sensing using microelectrodes made of monolayer graphene in a microfluidic device. The graphene microelectrodes exhibit improved electrolysis stability compared to traditional microelectrodes and allow for efficient separation and concentration of proteins. Based on the high optical transparency of graphene, the authors also developed a high-sensitivity three-dimensional multistream microfluidic strategy for label-free protein detection.
Review
Biochemical Research Methods
Lily Farmerie, Richard R. Rustandi, John W. Loughney, Mohamed Dawod
Summary: This review article highlights the significant recent advances in Isoelectric Focusing from 2015-2020, discussing principles, challenges, applications, and recent developments in various modes of Isoelectric Focusing on different platforms, as well as future directions for the technique.
JOURNAL OF CHROMATOGRAPHY A
(2021)
Article
Chemistry, Analytical
Jicheng Niu, Zhihui Bao, Zining Wei, Jasmine Xinze Li, Bin Gao, Xiaofan Jiang, Fei Li
Summary: A three-dimensional paper-based isoelectric focusing platform was developed for simultaneous separation and enrichment of proteins in both low-salt and high-salt samples. The platform showed efficient protein separation within 18 minutes and demonstrated good performance in pretreating high-salt samples, as well as accurately quantifying microalbuminuria and C-reactive proteins in clinical samples.
ANALYTICAL CHEMISTRY
(2021)
Review
Biochemical Research Methods
Gyula Vigh, Bohuslav Gas
Summary: In 1961, Svensson first described the principle of isoelectric focusing (IEF), which used a natural pH gradient formed by passing current through a sucrose density gradient-stabilized ampholyte mixture in a constant cross-section apparatus to separate ampholytic compounds. When polyacrylamide gel replaced the sucrose density gradient, the spatial and temporal stability of Svensson's pH gradient was lost, leading to a search for an explanation and improvement. It was later suggested that in the modern open-system IEF experiment, a two-stage process occurs, with a rapid first phase of linear pH gradient development followed by a slow second stage where isotachophoretic processes cause the pH gradient to decay as extreme pI CAs move into the electrode vessels.
Article
Multidisciplinary Sciences
Mehdi Rahmanian, Omid Sartipzadeh Hematabad, Esfandyar Askari, Farhad Shokati, Atin Bakhshi, Shiva Moghadam, Asiie Olfatbakhsh, Esmat Al Sadat Hashemi, Mohammad Khorsand Ahmadi, Seyed Morteza Naghib, Nidhi Sinha, Jurjen Tel, Hossein Eslami Amirabadi, Jaap M. J. den Toonder, Keivan Majidzadeh-A
Summary: This study demonstrates the use of a novel micropillar array-based microfluidic chip (MPA-Chip) for isolating circulating tumor cells (CTCs) from the blood of cancer patients, providing prognostic information and monitoring treatment efficacy. The lozenge geometry of the MPA-Chip was found to be the most effective design, with high capture efficiency (>85%), purity (>90%), and viability (97%).
JOURNAL OF ADVANCED RESEARCH
(2023)
Article
Chemistry, Analytical
Filip Dusa, Dana Moravcova, Karel Slais
Summary: We present a set of novel low-molecular-mass compounds with ampholytic properties that can be used as markers of isoelectric point in different isoelectric focusing formats. These compounds have excellent focusing properties and stability, making them suitable for various isoelectric focusing methods.
ANALYTICA CHIMICA ACTA
(2022)
Article
Multidisciplinary Sciences
Naotomo Tottori, Takasi Nisisako
Summary: Deterministic lateral displacement (DLD) is a promising technology for high-resolution separation of suspended particles based on size. Conventional DLD has a fixed critical diameter (D-c) determined by device geometry, but we propose a novel DLD that uses thermo-responsive hydrogel pillars to flexibly tune the D-c value. By adjusting temperature, the pillers can shrink or swell, allowing continuous switching of particle trajectories and on/off separation of particles.
SCIENTIFIC REPORTS
(2023)
Article
Biochemical Research Methods
Lin Huang, Qiang Zhang, Youli Tian, Xiaoping Liu, Weiwen Liu, Hua Xiao, Yuxin Wang, Chengxi Cao
Summary: An online array ultraviolet detection system was developed for real-time monitoring of protein fractions during Free-flow isoelectric focusing (FFIEF) process, showing good linearity and stability in experiments.
JOURNAL OF CHROMATOGRAPHY A
(2022)
Article
Biochemical Research Methods
Gyula Vigh, Bohuslav Gas
Summary: The carrier ampholytes-based isoelectric focusing has evolved from a closed system with steady pH gradient to an open system with transient pH gradient. The open system follows a two-stage model, where a rapid IEF process forms the pH gradient in the first stage, and isotachophoretic processes degrade the pH gradient in the second stage. The outcome of the process is determined by ionic mobilities, pK(a) values, and loaded amounts of all components, and a steady state cannot be achieved in terms of spatial coordinate.
Article
Chemistry, Analytical
Filip Dusa, Jiri Salplachta, Marie Horka, Kamila Lunerova, Katerina Rosenbergova, Oldrich Kubicek
Summary: We have developed a planar chip with a divergent geometry of separation channel for vertical free-flow electrophoresis. The chip has shown promising results in particle separation and analysis, especially for bacterial samples. This study has implications for the preparation and identification of bacteria using mass spectrometry.
ANALYTICA CHIMICA ACTA
(2022)
Review
Chemistry, Multidisciplinary
Tian Xu, Liangliang Sun
Summary: Capillary isoelectric focusing (cIEF) combined with electrospray ionization mass spectrometry (ESI-MS) and tandem mass spectrometry (MS/MS) is an ideal method for characterizing proteoforms based on their isoelectric points, but faces technical challenges that impede widespread adoption.
FRONTIERS IN CHEMISTRY
(2021)
Article
Biochemical Research Methods
Gyula Vigh, Bohuslav Gas
Summary: In modern isoelectric focusing systems, the formation of stationary, steady-state pH gradients is prevented due to convective mixing prevention, varying current densities, and imbalanced electrophoretic and diffusive fluxes. Open-system IEF is described as a two-stage process involving rapid IEF and subsequent degradation of the pH gradient. The authors propose a single process, transient, bidirectional isotachophoresis (tbdITP) model for open-system IEF in their previous paper, and this paper demonstrates the operational features of the tbdITP model using Simul 6.
Review
Chemistry, Analytical
Jiaqi Wu, Will McElroy, Janusz Pawliszyn, Christopher D. Heger
Summary: This article reviews the applications of icIEF technology in the pharmaceutical industry, highlighting the benefits of whole-column imaging detection for capillary isoelectric focusing. It also explores recent research using this technology to support advancements in the therapeutic landscape.
TRAC-TRENDS IN ANALYTICAL CHEMISTRY
(2022)
Article
Biotechnology & Applied Microbiology
Milad Radiom, Yong He, Juan Peng-Wang, Armelle Baeza-Squiban, Jean-Francois Berret, Yong Chen
BIOTECHNOLOGY AND BIOENGINEERING
(2020)
Article
Chemistry, Analytical
Zhenting Zhao, Yongjiao Sun, Jinxiang Song, Yajia Li, Yun Xie, Han Cui, Weiping Gong, Jie Hu, Yong Chen
Summary: A highly sensitive nonenzymetic electrochemical glucose sensor was developed using a multicomponent sensing electrode made of nickel cobalt layered double hydroxide and cobalt copper carbonate hydroxide on Cu foam. The sensor exhibited high sensitivity, large linear concentration range, and fast response times for glucose detection, making it applicable for micromole level glucose determination.
SENSORS AND ACTUATORS B-CHEMICAL
(2021)
Article
Nanoscience & Nanotechnology
Boxin Huang, Juan Peng, Xiaochen Huang, Feng Liang, Li Wang, Jian Shi, Ayako Yamada, Yong Chen
Summary: The research focuses on a method to generate interconnected neural clusters in a multiscale scaffold, leading to the formation of two types of neural clusters and indicating the complexity of neural-scaffold interaction and the variability of neural organization.
ACS APPLIED MATERIALS & INTERFACES
(2021)
Article
Engineering, Biomedical
Sisi Li, Momoko Yoshioka, Junjun Li, Li Liu, Shixin Ye, Ken-ichiro Kamei, Yong Chen
Summary: A casting method is developed to replicate the nanofibrous morphology onto a thin layer of PDMS, which proves to be a flexible and cost-effective approach for propagating hiPSCs.
BIOMEDICAL MATERIALS
(2022)
Article
Engineering, Biomedical
Yong He, Elrade Rofaani, Xiaochen Huang, Boxin Huang, Feng Liang, Li Wang, Jian Shi, Juan Peng, Yong Chen
Summary: A method using human induced pluripotent stem cells and artificial basement membrane to generate alveolar epithelium is presented. The alveolar epithelium achieved high-level expressions of specific proteins and characteristic tight junctions. Additionally, the method can potentially be applied to other types of artificial epithelium and endothelium.
ADVANCED HEALTHCARE MATERIALS
(2022)
Article
Engineering, Biomedical
Elrade Rofaani, Yong He, Juan Peng, Yong Chen
Summary: This study developed a method to overcome the lack of in vivo-like basement membrane (BM) for in vitro modeling by culturing primordial lung progenitors (PLPs) on an artificial basement membrane (ABM) composed of collagen IV and laminin. The forces generated by the growing epithelial monolayer on the ABM led to geometry-dependent folding and enhanced expression of surfactant protein C and aquaporin 5 compared with a nanofiber-covered substrate. This work emphasizes the importance of recapitulating natural BM for advanced epithelial modeling.
ACTA BIOMATERIALIA
(2023)
Article
Engineering, Biomedical
Wang Xi, Jad Saleh, Ayako Yamada, Caterina Tomba, Barbara Mercier, Sebastien Janel, Tien Dang, Matis Soleilhac, Aurelie Djemat, Huiqiong Wu, Beatrice Romagnolo, Frank Lafont, Rene-Marc Mege, Yong Chen, Delphine Delacour
Summary: This study developed hydrogel scaffolds with different elasticities and investigated their influence on the expansion, organization, and differentiation of intestinal epithelial cells. The results showed that cells on soft substrates exhibited improved characteristics resembling in vivo intestinal tissue, which is of significant importance for the design of biomaterials for ex vivo intestinal models.
Article
Biochemistry & Molecular Biology
Elrade Rofaani, Boxin Huang, Feng Liang, Juan Peng, Yong Chen
Summary: This study successfully generated airway epithelium using artificial basement membrane (ABM) and human-induced pluripotent stem cells (hiPSCs). The generated pseudostratified airway epithelium consists of multiple cell types and demonstrates advantages over other methods.
INTERNATIONAL JOURNAL OF BIOLOGICAL MACROMOLECULES
(2022)
Article
Materials Science, Biomaterials
Xiaochen Huang, Boxin Huang, Yong He, Liang Feng, Jian Shi, Li Wang, Juan Peng, Yong Chen
Summary: SARS-CoV-2 may cause molecular and functional changes in cardiomyocytes through the interaction with ACE2 complex. The spike proteins of SARS-CoV-2 can lead to abnormal sarcomere structures and disrupted gap junction protein distribution in human-induced pluripotent stem cell-derived cardiomyocytes. Additionally, the damage to cardiomyocytes caused by spike proteins is enhanced when angiotensin II is added, resulting in potential cardiac dysfunction.
Article
Engineering, Biomedical
Boxin Huang, Yong He, Elrade Rofaani, Feng Liang, Xiaochen Huang, Jian Shi, Li Wang, Ayako Yamada, Juan Peng, Yong Chen
Summary: The study introduced a method for directed differentiation of hiPSCs using nanofiber membranes, resulting in the formation of regular and interconnected cortical neural clusters. By modulating the substrate stiffness and permeability, cells were concentrated in the central area of the honeycomb compartments, with synchronized neural activities. Compared to other types of substrates, nanofiber membranes were more efficient in promoting cell self-organization and neural activities.
ACTA BIOMATERIALIA
(2022)
Article
Materials Science, Biomaterials
Changchong Chen, Zeinab Ibrahim, Marion F. Marchand, Tristan Piolot, Sahil Kamboj, Franck Carreiras, Ayako Yamada, Marie-Claire Schanne-Klein, Yong Chen, Ambroise Lambert, Carole Aime
Summary: Cellular heterogeneity plays a critical role in various physiological processes, and the extracellular matrix (ECM) is crucial for generating this heterogeneity. The researchers have developed ECM models that allow independent control of protein composition, topology, and stiffness, which can recreate the complex phenotypic landscape of ovarian cancer cells.
ACS BIOMATERIALS SCIENCE & ENGINEERING
(2022)
Article
Nanoscience & Nanotechnology
Feng Tian, Linlin Yin, Peiran Lin, Yurong Liu, Wenlong Wang, Yong Chen, Yadong Tang
Summary: In this study, a micro-/nanohierarchical substrate was fabricated using soft lithography and electrospinning to evaluate the synergistic effect of aligned nanofibrous topographical guidance and off-ground culture environment on the formation of endothelium and maturation of hiPSC-CMs. The study showed that the micro-/nanohierarchical substrate enhanced the proliferation, alignment, and cell-cell interaction of HUVECs, as well as the sarcomere length and maturation-related gene expression of hiPSC-CMs. Moreover, hiPSC-CMs on the substrate displayed higher resistance to cardiac drugs compared to other groups, indicating higher maturity. Overall, the proposed substrate holds great potential for in vitro models and tissue engineering.
ACS APPLIED MATERIALS & INTERFACES
(2023)
Article
Biophysics
Xiaochen Huang, Feng Liang, Boxin Huang, Haoyue Luo, Jian Shi, Li Wang, Juan Peng, Yong Chen
Summary: Recent advances have shown that electrochemical impedance spectroscopy is highly sensitive in real-time monitoring of cell barriers on a chip. In this study, the method was applied to investigate the hiPSC derived and ABM-supported endothelial barrier. The results showed enhanced stability of the BMEC barrier in a serum-free and BrdU-containing culture medium compared to conventional culture, and the barrier was sensitive to stimuli such as thrombin. This method can be advocated for investigating cell barrier integrity and barrier-based assays.
BIOSENSORS & BIOELECTRONICS
(2023)
Article
Engineering, Electrical & Electronic
Changchong Chen, Yong He, Elliot Lopez, Franck Carreiras, Ayako Yamada, Marie-Claire Schanne-Klein, Ambroise Lambert, Yong Chen, Carole Aime
Summary: We developed an invasion assay using microfabricated culture devices that can control the number and size of spheroids. We observed spheroid disruption due to substrate-induced cell migration. This method holds potential for tumor modeling and anti-cancer drug development.
MICRO AND NANO ENGINEERING
(2022)
Article
Engineering, Electrical & Electronic
Feng Liang, Juan Peng, Jian Shi, Li Wang, Carole Aime, Yong Chen
Summary: This study presents a method utilizing a microfluidic device to capture and release rare cells. By applying reverse flow and adding micro particles, the efficiency of capture and release can be improved.
MICRO AND NANO ENGINEERING
(2022)
