Bioorthogonal Microbubbles with Antifouling Nanofilm for Instant and Suspended Enrichment of Circulating Tumor Cells

Abstract

Integrating clinical rare cell enrichment, culture, and single-cell phenotypic profiling is currently hampered by the lack of competent technologies, which typically suffer from weak cell−interface collision affinity, strong nonspecific adsorption, and the potential uptake. Here, we report cellson-a-bubble, a bioinspired, self-powered bioorthogonal microbubble (click bubble) that leverages a clickable antifouling nanointerface and a DNA-assembled sucker-like polyvalent cell surface, to enable instant and suspended isolation of circulating tumor cells (CTCs) within minutes. Using this biomimetic engineering strategy, click bubbles achieve a capture efficiency of up to 98%, improved by 20% at 15 times faster over their monovalent counterparts. Further, the buoyancy-activated bubble facilitates self-separation, 3D suspension culture, and in situ phenotyping of the captured single cancer cells. By using a multiantibody design, this fast, affordable micromotor-like click bubble enables suspended enrichment of CTCs in a cohort (n = 42) across three cancer types and treatment response evaluation, signifying its great potential to enable single-cell analysis and 3D organoid culture.

Buoyant Metal–Organic Framework Corona-Driven Fast Isolation and Ultrasensitive Profiling of Circulating Extracellular Vesicles

“…To achieve rapid isolation of EVs, we engineered a buoyancy-driven, EV-affinity coordination corona by assembling a Zr-MOF on the surface of a floating glass bubble (∼15 μm in diameter), inspired by the strong adhesion of barnacles on a swimming whale. As shown in Figure a, we first coated a polyethylenimine (PEI) nanofilm on the bubble surface via electrostatic assembly.…”

CRISPR/dCas9-Mediated Specific Molecular Assembly Facilitates Genotyping of Mutant Circulating Tumor DNA

“…Taking these advantages, multiple recognition units could be designed in an array to achieve simultaneous genotyping of different ctDNA targets. , The signal output system can also be integrated with other signal amplification methods, such as DNA assembly, strand displacement, and metal–organic framework caging, ,− to realize ultrasensitive ctDNA detection. We conceive that our E-dCas9 design could be further combined with microfluidics and functional nanostructures, − especially PAM-free Cas9, for sensitive, specific, and multiplexed detection of ctDNA on a fully integrated diagnostic platform to better meet practical needs in clinical settings.…”

A Novel Multiarmed Bifunctional PEG Derivative for the Preparation of Mass Spectrometry Ion Sources with Antifouling Property and High Selectivity

“…Recently, the click reaction has rapidly become a powerful tool for the immobilization of biomolecules due to its strong selectivity, high efficiency, and mild reaction condition . The copper-free reaction of trans -cycloalkene (TCO) with 1,2,4,5-tetrazine (Tz) has a rate constant of 3.3 × 10 6 M –1 s –1 (in water at 25 °C), which is one of the fastest bio-orthogonal reactions known to date. , Therefore, to prepare an MS ionization substrate with antifouling properties, high selectivity, and sensitivity, it is beneficial to use polydopamine as the connection medium between the PEG derivative and substrate and click chemistry as the immobilization strategy of biomolecules.…”