Visible photorelease of liquid biopsy markers following microfluidic affinity-enrichment

Pahattuge, Thilanga N.; Jackson, Joshua M.; Rane, Digamber; Wijerathne, Harshani; Brown, Virginia; Witek, Małgorzata A.; Perera, Chamani; Givens, Richard S.; Peterson, Blake R.

doi:10.1039/c9cc09598e

Cited by 13 publications

(28 citation statements)

References 22 publications

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“…The attractive nature of this design is that it can process whole blood directly yielding high recovery of CTCs with high purity . The selection module was replicated in COC because this material has excellent optical properties that enable efficient UV/O 3 activation of the high-aspect-ratio microchannels for high loads of antibody onto the activated channels using the PC linker as well as high transmissivity in the blue region of the electromagnetic spectrum to allow for efficient CTC release …”

Section: Resultsmentioning

confidence: 91%

“…Modules were connected to the motherboard, and the SMART-Chip was washed with 0.5% BSA/PBS (0.1 mL, 20 μL/min) prior to blood infusion. Stained cells were spiked into healthy blood and hydrodynamically introduced (1 mL, 25 μL/min) into the CTC selection module that was surface-decorated with anti-EpCAM monoclonal antibodies via a photocleavable (PC) linker . Following blood infusion, the module was rinsed with 1 mL of 0.5% BSA/PBS (50 μL/min) to remove any nonspecifically bound material.…”

Section: Methodsmentioning

confidence: 99%

“…Three task-specific modules consisted of a CTC enrichment module, impedance module, and imaging module. The SMART-Chip could perform the following steps: (i) affinity-select CTCs; (ii) release of the enriched biomarkers using a photocleavable (PC) linker; (iii) counting of the released cells using an impedance sensor that also determined cell viability; and (iv) immunophenotyping …”

mentioning

confidence: 99%

“…Three taskspecific modules consisted of a CTC enrichment module, impedance module, and imaging module. The SMART-Chip could perform the following steps: (i) affinity-select CTCs; (ii) release of the enriched biomarkers using a photocleavable (PC) linker; 38 (iii) counting of the released cells using an impedance sensor that also determined cell viability; and (iv) immunophenotyping. 39 The CTC enrichment module consisted of an array of highaspect-ratio sinusoidally shaped microchannels arranged in a zconfiguration 40 and provided high cell recovery (>80%) with exquisite purity (>85%).…”

mentioning

confidence: 99%

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System Modularity Chip for Analysis of Rare Targets (SMART-Chip): Liquid Biopsy Samples

et al. 2021

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Liquid biopsies are becoming popular for managing a variety of diseases due to the minimally invasive nature of their acquisition, thus potentially providing better outcomes for patients. Circulating tumor cells (CTCs) are among the many different biomarkers secured from a liquid biopsy, and a number of efficient platforms for their isolation and enrichment from blood have been reported. However, many of these platforms require manual sample handling, which can generate difficulties when translating CTC assays into the clinic due to potential sample loss, contamination, and the need for highly specialized operators. We report a system modularity chip for the analysis of rare targets (SMART-Chip) composed of three taskspecific modules that can fully automate processing of CTCs. The modules were used for affinity selection of the CTCs from peripheral blood with subsequent photorelease, simultaneous counting, and viability determinations of the CTCs and staining/imaging of the CTCs for immunophenotyping. The modules were interconnected to a fluidic motherboard populated with valves, interconnects, pneumatic control channels, and a fluidic network. The SMART-Chip components were made from thermoplastics via microreplication, which lowers the cost of production making it amenable to clinical implementation. The utility of the SMART-Chip was demonstrated by processing blood samples secured from colorectal cancer (CRC) and pancreatic ductal adenocarcinoma (PDAC) patients. We were able to affinity-select EpCAM expressing CTCs with high purity (0−3 white blood cells/mL of blood), enumerate the selected cells, determine their viability, and immunophenotype the cells. The assay could be completed in <4 h, while manual processing required >8 h.

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Section: Resultsmentioning

confidence: 91%

Section: Methodsmentioning

confidence: 99%

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confidence: 99%

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confidence: 99%

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System Modularity Chip for Analysis of Rare Targets (SMART-Chip): Liquid Biopsy Samples

et al. 2021

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“…Soper and coworkers (41,42) have demonstrated that surface immobilized (coumarin-4-yl)methyl as photocleavable linker 42 can be used to capture and enrich clinically relevant liquid biopsy markers, that is, circulating tumor cells and extracellular vesicles, with an affinity agent such as an antibody, that is covalently attached to the photocleavable linker via its amino group, Scheme 10. Irradiation with blue LED light (412 nm) is used to cleave the long chain ester from the C-4 methyl of the coumarin to release the Ab-CTC or Ab-EV complexes in > 90% yields.…”

Section: Representative Applicationsmentioning

confidence: 99%

The Discovery, Development and Demonstration of Three Caged Compounds^†

Steinmetz

Givens

2021

Photochem & Photobiology

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An overview of the history, mechanistic aspects and applications is provided for p-hydroxyphenacyl (pHP) and benzoin photoremovable protecting groups, which release biologically important leaving groups upon photolysis with UV light. Also discussed is (7-diethylaminocoumarin-4-yl)methyl (DEACM), a photoremovable protecting group that absorbs visible light. These are followed by the a-keto amides and naphtho-and benzothiophene-2-carboxanilides as caging groups, which eliminate leaving groups via photochemically produced zwitterionic intermediates. Also covered are amino-1,4benzoquinones, which upon exposure to green and red wavelengths of light photorearrange to an unstable photoproduct that subsequently eliminates leaving groups in aqueous media. Selected examples are given that use these photoremovable protecting (caging) groups for the light-activated release of biologically important substrates under physiological conditions in cells and tissue as practical applications in biology, biochemistry and physiology. These caging groups have found significant applications because their photochemistry is efficient and a single coproduct is formed in addition to the photoreleased substrate.

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Aryl‐Diazonium Salts Offer a Rapid and Cost‐Efficient Method to Functionalize Plastic Microfluidic Devices for Increased Immunoaffinity Capture

Rabe,

Ho,

Choudhury

et al. 2023

Adv Materials Technologies

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Microfluidic devices have been used for decades to isolate cells, viruses, and proteins using on‐chip immunoaffinity capture using biotinylated antibodies, proteins, or aptamers. To accomplish this, the inner surface is modified to present binding moieties for the desired analyte. While this approach is successful in research settings, it is challenging to scale many surface modification strategies. Traditional polydimethylsiloxane (PDMS) devices can be effectively functionalized using silane‐based methods; however, it requires high labor hours, equipment, and hazardous chemicals. Manufacture of microfluidic devices using plastics, including cyclic olefin copolymer (COC), allows chips to be mass produced, but most functionalization methods used with PDMS are not compatible with plastic. Herein, this work demonstrates how to deposit biotin onto the surface of a plastic microfluidic chips using aryl‐diazonium. This method chemically bonds biotin to the surface, allowing for the addition of streptavidin nanoparticles to the surface. Nanoparticles increase the surface area of the chip and allow for proper capture moiety orientation. This process is faster, can be performed outside of a fume hood, is very cost‐effective using readily available laboratory equipment, and demonstrates higher rates of capture. Additionally, this method allows for more rapid and scalable production of devices, including for diagnostic testing.

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Visible photorelease of liquid biopsy markers following microfluidic affinity-enrichment

Abstract: We detail a heterobifunctional, 7-aminocoumarin photocleavable (PC) linker with unique properties to covalently attach Abs to surfaces and subsequently release them with visible light (400–450 nm).

Cited by 13 publications

References 22 publications

System Modularity Chip for Analysis of Rare Targets (SMART-Chip): Liquid Biopsy Samples

System Modularity Chip for Analysis of Rare Targets (SMART-Chip): Liquid Biopsy Samples

The Discovery, Development and Demonstration of Three Caged Compounds^†

Aryl‐Diazonium Salts Offer a Rapid and Cost‐Efficient Method to Functionalize Plastic Microfluidic Devices for Increased Immunoaffinity Capture

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Visible photorelease of liquid biopsy markers following microfluidic affinity-enrichment

Abstract: We detail a heterobifunctional, 7-aminocoumarin photocleavable (PC) linker with unique properties to covalently attach Abs to surfaces and subsequently release them with visible light (400–450 nm).

Cited by 13 publications

References 22 publications

System Modularity Chip for Analysis of Rare Targets (SMART-Chip): Liquid Biopsy Samples

System Modularity Chip for Analysis of Rare Targets (SMART-Chip): Liquid Biopsy Samples

The Discovery, Development and Demonstration of Three Caged Compounds†

Aryl‐Diazonium Salts Offer a Rapid and Cost‐Efficient Method to Functionalize Plastic Microfluidic Devices for Increased Immunoaffinity Capture

Contact Info

Product

Resources

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The Discovery, Development and Demonstration of Three Caged Compounds^†