Combinatorial Immunophenotyping of Cell Populations with an Electronic Antibody Microarray

Chu, Chia‐Heng; Wang, Ningquan; Ozkaya‐Ahmadov, Tevhide; Civelekoglu, Ozgun; Lee, Dohwan; Arifuzzman, A K M; Sarioglu, A. Fatih

doi:10.1002/smll.201904732

Cited by 15 publications

(12 citation statements)

References 45 publications

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“…22−24 In addition, several hurdles limit the application of the highly experience dependent technique, such as inevitable measurements errors, requirement of an extremely expensive instrument, and high-content data analysis. 25 Comparably, we report a simple, economic, and efficient alternative approach, an electrochemical phenotyping method, in this paper, that facilitates reliable, rapid, and low-cost characterization of stemlike phenotypes in breast cancer. To present the expression signatures of the stemlike biomarkers, we incorporated a DNA assembly machinein situ programmable DNA circuit (PDC)to construct topologically defined nanostructures at the biomolecular recognition interface, and permits ultrasensitive phenotypic assay by translating the target binding events to the quantitative signals from well-organized strand crossover events.…”

Section: ■ Introductionmentioning

confidence: 95%

“…As such, compensation at the detectors and “gating” are always necessary to correct the signal spillover and outline the distribution regions of cell subpopulations. However, incorrect compensation could adversely affect the sensitivity and lead to the generation of unexpected cell subpopulations; there is a “gating” effect wherein a lack of standardized instrument setup requires reliance exclusively on the people with experience. − In addition, several hurdles limit the application of the highly experience dependent technique, such as inevitable measurements errors, requirement of an extremely expensive instrument, and high-content data analysis . Comparably, we report a simple, economic, and efficient alternative approach, an electrochemical phenotyping method, in this paper, that facilitates reliable, rapid, and low-cost characterization of stemlike phenotypes in breast cancer.…”

Section: Introductionmentioning

confidence: 96%

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In Situ Programmable DNA Circuit-Promoted Electrochemical Characterization of Stemlike Phenotype in Breast Cancer

Cao

Han

et al. 2021

J. Am. Chem. Soc.

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Breast cancer is one of the most common malignant diseases among women worldwide, and the existence of breast cancer stem cells is closely associated with poor outcomes. Herein, we report an electrochemical phenotyping method to characterize the stemlike phenotype in breast cancer, offering a low-cost but robust choice other than the highly expensive and experience-dependent flow cytometry. Specially, after immune-magnetic beads-assisted enrichment, an in situ programmable DNA circuit is designed using capture probes to bring in the toeholds for DNA assembly and effector probes to accelerate the removal of background signals. The electrochemical phenotyping method could sensitively determine breast cancer stem cells in a wide linear range and exhibit desirable accuracy and reliability. The method can not only monitor the phenotypic transition of breast cancer cells and the drug-reversed effect but also determinate stemlike phenotype in the mice bearing breast cancer xenograft tumor. Overall, the electrochemical phenotyping method may provide promising technical support for precise management of breast tumors.

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Section: ■ Introductionmentioning

confidence: 95%

Section: Introductionmentioning

confidence: 96%

In Situ Programmable DNA Circuit-Promoted Electrochemical Characterization of Stemlike Phenotype in Breast Cancer

Cao

Han

et al. 2021

J. Am. Chem. Soc.

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“…and other researchers developed microfluidic platforms for electrophoresis and sample separation purposes at the beginning of the 1990s (Manz et al., 1990 ; Harrison et al., 1992 ; Mathies & Huang, 1992 ; Woolley & Mathies, 1994 ). Microfluidics was then used as a multidisciplinary breakthrough technology that was quickly established with wide applications as in environmental sensing (Vasudev et al., 2013 ; Pol et al., 2017 ), combinatorial chemistry (synthesis and assays) (Li et al., 2018 ; Liu et al., 2019 ), energy applications (Chen et al., 2018 ; De et al., 2020 ), micro-propulsion (Feng et al., 2015 ; Serrano et al., 2018 ), organ-on-a-chip technology (Sittadjody et al., 2021 ; Zheng et al., 2021 ), clinical diagnostics (Luo et al., 2005 ; Chango et al., 2006 ), and drug delivery (Bains et al., 2017 ; Soheili et al., 2021 ). This review focuses on the application of microfluidics in drug delivery.…”

Section: Introductionmentioning

confidence: 99%

Merits and advances of microfluidics in the pharmaceutical field: design technologies and future prospects

et al. 2022

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Microfluidics is used to manipulate fluid flow in micro-channels to fabricate drug delivery vesicles in a uniform tunable size. Thanks to their designs, microfluidic technology provides an alternative and versatile platform over traditional formulation methods of nanoparticles. Understanding the factors that affect the formulation of nanoparticles can guide the proper selection of microfluidic design and the operating parameters aiming at producing nanoparticles with reproducible properties. This review introduces the microfluidic systems’ continuous flow (single-phase) and segmented flow (multiphase) and their different mixing parameters and mechanisms. Furthermore, microfluidic approaches for efficient production of nanoparticles as surface modification, anti-fouling, and post-microfluidic treatment are summarized. The review sheds light on the used microfluidic systems and operation parameters applied to prepare and fine-tune nanoparticles like lipid, poly(lactic-co-glycolic acid) (PLGA)-based nanoparticles as well as cross-linked nanoparticles. The approaches for scale-up production using microfluidics for clinical or industrial use are also highlighted. Furthermore, the use of microfluidics in preparing novel micro/nanofluidic drug delivery systems is presented. In conclusion, the characteristic vital features of microfluidics offer the ability to develop precise and efficient drug delivery nanoparticles.

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“…Based on these studies, an antibody microarray was successfully applied for the quality control of mesenchymal stem cells . Other research groups also reported the potential use of similar antibody microarrays in immunophenotyping. − …”

Section: Introductionmentioning

confidence: 99%

Quantitative Cell Subset Analysis Using Antibody Microarrays

Ogasawara

Kuwabara

Kozai

et al. 2021

ACS Appl. Bio Mater.

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The expression patterns of surface antigens are associated with the differentiation status and functional characteristics of mammalian cells. To analyze the surface antigen expression pattern in a high-throughput manner, antibody microarrays have been developed by several groups, including ours. This analysis can be performed using cell-binding assays on microarrays; moreover, this approach has advantages over conventional flow cytometry (FCM). Unlike FCM, the microarray-based method cannot evaluate the concurrent expression of more than two surface antigens on a single cell, and therefore, it cannot be used for cell subset analysis. To overcome this drawback, we prepared an antibody microarray with spots presenting co-immobilized multiple antibodies together with spots presenting each antibody separately. The co-immobilized spots are expected to be reactive for every surface antigen specific to the co-immobilized antibodies. In addition, the concept of an algebra of sets is incorporated into the derivation of quantitative data regarding cell subsets. Here, taking cell subsets with respect to two surface antigens as the simplest example, antibody microarrays were prepared and initially subjected to validation studies to verify the accuracy of cell-binding assays. Quantitative subset analysis was performed using antibody microarrays prepared using the anti-CD13 and anti-CD49f antibodies. For model populations that consisted of discrete subsets, THP-1, HL-60, CCRF-CEM, and Ramos cell lines were used because they were found by FCM to have a singular phenotype, that is, CD13+CD49f+, CD13+CD49f–, CD13–CD49f+, and CD13–CD49f–, respectively. Five populations were prepared by mixing these cells at various ratios and analyzed for their subsets using microarrays. The results showed that the experimentally determined abundance ratios of the four model subsets were in good agreement with the predetermined abundance ratios, which provided the proof of principle for the new method in the quantitative subset analysis.

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Combinatorial Immunophenotyping of Cell Populations with an Electronic Antibody Microarray

Abstract: presents opportunities in global health and telemedicine applications for cell-based diagnostics and health monitoring.

Cited by 15 publications

References 45 publications

In Situ Programmable DNA Circuit-Promoted Electrochemical Characterization of Stemlike Phenotype in Breast Cancer

In Situ Programmable DNA Circuit-Promoted Electrochemical Characterization of Stemlike Phenotype in Breast Cancer

Merits and advances of microfluidics in the pharmaceutical field: design technologies and future prospects

Quantitative Cell Subset Analysis Using Antibody Microarrays

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