The study of exosomes has great significance for cancer diagnosis and prognosis, in which damage-free isolation of exosomes is a critically important step due to their extremely low concentration in body fluids. Herein, immunoaffinitive superparamagnetic nanoparticles (IS-NPs) were successfully constructed by combining antibodies with superparamagnetic nanoparticles through host–guest interactions between β-cyclodextrin (β-CD) and 4-aminoazobenzene (AAB). Exosomes in cell-culture supernatants or body fluids can be efficiently captured by IS-NPs and then mildly eluted by competitive host molecules, α-CD. Approximately 80% of the exosomes were captured by the IS-NPs from the model samples containing varied concentrations of exosomes (1011–107 exosomes per mL). After elution, the release efficiency of the captured exosomes was as high as 86.5%, as calculated from the extracted RNA content. A comparison between the exosomes isolated by our IS-NPs, conventional ultracentrifugation (UC), polyethylene glycol (PEG)-based precipitation, and a commercial kit was conducted to examine the performance of the IS-NP exosome isolation. The particle-to-protein ratio of IS-NP captured exosomes (8.8 ± 1.3 × 109) was eight-fold higher than that of the UC (1.1 ± 0.4 × 109) and more than two-fold that of the PEG-based precipitation (3.7 ± 0.8 × 109) and commercial kit (3.2 ± 0.8 × 109) isolated ones, revealing that IS-NPs yielded more exosomes with higher purity than all the other three approaches. Cellular uptake experiments demonstrated that the exosomes captured by IS-NPs have retained their structural and functional integrities, and their broad applications in biomedical areas could therefore be expected. Promisingly, this host–guest interaction involving immunoaffinity magnetic particle system could offer a new way for fast, high-efficiency and high-purity exosome isolation and elution, further benefitting exosomes’ applications in desired areas.
Magnetic guanidyl-functionalized metal–organic framework (MOF) nanospheres with multi-affinity sites (inherent Zn–O cluster, amino group, and guanidyl group) for efficient enrichment of global phosphopeptides.
Novel magnetic metal–organic framework nanocomposites with 1,4-phenylenebisboronic acid as both an organic ligand and a functional molecule are proposed for the first time as a new type of intelligent nanomaterial to selectively capture and release glycoproteins via pH-stimulus-response, and would be of great potential for use in bio-separation.
7 September 2022 in Journal of Colloid and Interface Science
Highly efficient isolation and recovery of viable circulating tumor cells (CTCs) from the blood of patients is an important precondition to address the current dilemma of insufficient CTC studies, and can promote the development of individualized antitumor therapies. Herein, a cell-friendly CTC isolation and recovery nanoplatform with luminescent labelling was established using a layer-by-layer (LbL) assembly technique and a stimulated cellular-release strategy. In particular, the anti-epithelial cell adhesion molecule (anti-EpCAM) antibody was introduced with a disulfide bond-containing linker for further bio-friendly recovery of the CTCs. Quantum dots (QDs) were deposited onto fast magnet-responsive Fe3O4 nanoparticles through a facile LbL assembly method to monitor the capture and recovery process in real time. The obtained PEGlyated immuno-magnetic nanospheres (PIMNs) can all be magnetically collected within 2 min. Capture efficiencies above 90% can be achieved from blood samples with 5–200 CTCs per mL after only 1–2 min incubation. Nearly all PIMNs on the surface of the CTCs were detached after 15 min of glutathione (GSH) treatment with the disappearance of QD signals. Recovered CTCs could be directly used for culture (cell viability, ∼98%), and their invasiveness and migration characteristics remained unchanged. Furthermore, the PIMNs were successfully applied to isolate CTCs in cancer patients’ peripheral blood samples, and an average of 8.6 ± 5.8 CTCs per mL was detected. The results above suggested that PIMNs may serve as a powerful nanoplatform for CTC screening, isolation and recovery.