RESUMO
To impart biomedical functions to nanoparticles (NPs), the common approach is to conjugate functional groups onto NPs by dint of the functions of those groups per se. It is still beyond current reach to create protein-like specific interactions and functions on NPs by conformational engineering of nonfunctional groups on NPs. Here, we develop a conformational engineering method to create an NP-based artificial antibody, denoted "Goldbody," through conformational reconstruction of the complementary-determining regions (CDRs) of natural antibodies on gold NPs (AuNPs). The seemingly insurmountable task of controlling the conformation of the CDR loops, which are flexible and nonfunctional in the free form, was accomplished unexpectedly in a simple way. Upon anchoring both terminals of the free CDR loops on AuNPs, we managed to reconstruct the "active" conformation of the CDR loops by tuning the span between the two terminals and, as a result, the original specificity was successfully reconstructed on the AuNPs. Two Goldbodies have been created by this strategy to specifically bind with hen egg white lysozyme and epidermal growth factor receptor, with apparent affinities several orders of magnitude stronger than that of the original natural antibodies. Our work demonstrates that it is possible to create protein-like functions on NPs in a protein-like way, namely by tuning flexible surface groups to the correct conformation. Given the apparent merits, including good stability, of Goldbodies, we anticipate that a category of Goldbodies could be created to target different antigens and thus used as substitutes for natural antibodies in various applications.
Assuntos
Anticorpos Monoclonais/química , Regiões Determinantes de Complementaridade/química , Ouro/química , Nanopartículas Metálicas/química , Conformação ProteicaRESUMO
The size effect on the cellular uptake of nanoparticles (NPs) has been extensively studied, but it is still not well understood. Herein, a reductionist approach is used to minimize all influencing factors except the particle size, and co-exposure of different-sized silica nanoparticles (SNPs) is adopted instead of the common single exposure. SNPs are found being internalized by Hela cells in serum-free medium mainly via clathrin-dependent endocytosis, thus simplifying the data analysis for reliable attribution to size effects. Remarkably, even though at conditions that the size effects seem very small or even undetectable in the common single exposure experiments, the co-exposure experiments reveal significant size effects due to an unexpected interplay between two different-sized SNPs. Namely, the bigger SNPs significantly promote the cellular uptake of the smaller ones, while the smaller SNPs inhibit the internalization of the bigger ones, with a total uptake increase of the particle number of SNPs in the cells. This strong interplay between different-sized NPs might unavoidably exist within most "single-sized" NP products, whose sizes actually distribute in certain ranges, thus urging reconsideration of the size effect on the cellular uptake of NPs, for the benefits of both bioapplications and safety assessment of nanomaterials.
Assuntos
Nanopartículas/química , Nanopartículas/metabolismo , Dióxido de Silício/química , Células HeLa , HumanosRESUMO
Carbon dots (CDots) in a general structure of small carbon nanoparticles with various surface passivation schemes have emerged to represent a new class of carbon nanomaterials in now a rapidly advancing and expanding research field. Among various synthesis methods, the use of pre-processed and selected small carbon nanoparticles for deliberate chemical functionalization of the particle surface with organic molecules have produced high-performance and structurally better defined CDots. Specifically, small organic molecules 2,2'-(ethylenedioxy)bis(ethylamine) and 3-ethoxypropylamine were used for the effective surface passivation of the carbon nanoparticles via chemical functionalization to yield CDots that are brightly fluorescent and also structurally ultra-compact, amenable to various desired cell imaging applications. Thus, a systematic evaluation of these CDots on their cytotoxicity profiles is necessary, and performed in this study by using a diverse selection of cell lines. Also for fluorescence imaging, CDots were modified with their encapsulating selected organic dyes for much enhanced red/near-IR fluorescence emissions. These modified CDots with the dyes as guest were also evaluated for their cytotoxicity profiles. The results suggest that the CDots without and with the guest dyes are generally nontoxic to the selected cell lines, further supporting the notion that CDots can be used as high-performance yet nontoxic bioimaging agents.
Assuntos
Nanoestruturas , Pontos Quânticos , Carbono/toxicidade , Corantes , Imagem Óptica , Pontos Quânticos/toxicidadeRESUMO
Carbon dots (CDots) for their excellent optical and other properties have been widely pursued for potential biomedical applications, in which a more comprehensive understanding on the cellular behaviors and mechanisms of CDots is required. For such a purpose, two kinds of CDots with surface passivation by 3-ethoxypropylamine (EPA-CDots) and oligomeric polyethylenimine (PEI-CDots) were selected for evaluations on their uptakes by human cervical carcinoma HeLa cells at three cell cycle phases (G0/G1, S and G2/M), and on their different internalization pathways and translocations in cells. The results show that HeLa cells could internalize both CDots by different pathways, with an overall slightly higher internalization efficiency for PEI-CDots. The presence of serum in culture media could have major effects, significantly enhancing the cellular uptake of EPA-CDots, yet markedly inhibiting that of PEI-CDots. The HeLa cells at different cell cycle phases have different behaviors in taking up the CDots, which are also affected by the different dot surface moieties and serum in culture media. Mechanistic implications of the results and the opportunities associated with an improved understanding on the cellular behaviors of CDots for potentially the manipulation and control of their cellular uptakes and translocations are discussed.
Assuntos
Carbono/farmacocinética , Pontos Quânticos/química , Carbono/química , Sobrevivência Celular/efeitos dos fármacos , Células HeLa , Humanos , Tamanho da Partícula , Polietilenoimina/química , Polietilenoimina/farmacocinética , Propilaminas/química , Propilaminas/farmacocinética , Propriedades de Superfície , Células Tumorais CultivadasRESUMO
Exploring novel drug delivery systems with good stability and new structure to integrate pillararene and upconversion nanoparticles (UCNPs) into one system continues to be an important challenge. Herein, we report a novel preparation of a supramolecular upconversion nanosystem via the host-guest complexation based on carboxylate-based pillar[5]arene (WP5) and 15-carboxy-N,N,N-trialkylpentadecan-1-ammonium bromide (1)-functionalized UCNPs to produce WP5â1-UCNPs that can be loaded with the chemotherapeutic drug doxorubicin (DOX). Importantly, the WP5 on the surface of the drug-loaded nanosystem can be efficiently protonated under acidic conditions, resulting in the collapse of the nanosystem and drug release. Moreover, cellular uptake confirms that the nanosystem can enter human cervical cancer (HeLa) cells, resulting in drug accumulation in the cells. More importantly, cytotoxicity experiments demonstrated the excellent biocompatibility of WP5â1-UCNPs without loading DOX and that the nanosystem DOX-WP5â1-UCNPs exhibited an ability of killing HeLa cells effectively. We also investigated magnetic resonance imaging and upconversion luminescence imaging, which may be employed as visual imaging agents in cancer diagnosis and treatment. Thus, in the present work, we show a simple yet powerful strategy to combine UCNPs and pillar[5]arene to produce a unified nanosystem for dual-mode bioimaging-guided therapeutic applications.