RESUMO
With the advancement of life medicine, in vitro diagnostics (IVD) technology has become an auxiliary tool for early diagnosis of diseases. However, biosensors for IVD now face some disadvantages such as poor targeting, significant antifouling properties, low density of recognized molecules, and poor stability. In recent years, peptides have been demonstrated to have various functions in unnatural biological systems, such as targeting properties, antifouling properties, and self-assembly properties, which indicates that peptides can be engineered. These properties of peptides, combined with their good biocompatibility, can be well applied to the design of biosensors to solve the problems mentioned above. This review provides an overview of the properties of engineered functional peptides and their applications in enhancing biosensor performance, mainly in the field of optics and electrochemistry.
Assuntos
Técnicas Biossensoriais , Peptídeos , EletroquímicaRESUMO
Biosensors developed through a sandwich approach have demonstrated favorable detection performance for exosomal programmed cell death 1 ligand 1 (ExoPD-L1) detection. However, the reported PD-L1 antibodies, peptides, and aptamers utilized in these biosensors typically bind to the extracellular region, with overlapping binding sites that severely constrain the fabrication of biosensors. In this study, we present a simple approach to specifically identify and analyze ExoPD-L1 through the non-selective trapping effect of Ti3C2TX (X=-O, -F, -OH) MXene on exosomes via the formation of Ti-O-P complexation, and the selective capture of peptide-functionalized Au@MPBA (4-Mercaptophenylboronic acid) @SiO2 surface enhanced Raman scattering (SERS) tags on ExoPD-L1. The biosensor delivered a both hypersensitive and reliable performance in exosome detection with a low limit of detection (20.74 particles/mL) in the linear range of 102 to 5×106 particles/mL. Furthermore, the biosensor demonstrated excellent stability and interference resistance in detecting ExoPD-L1 in clinical serum samples, enabling the easy differentiation of breast cancer patients from healthy controls. This work provides new insights into the design of biosensors for exosome detection and can serve as a replicable template for sandwich immunoassay detection for other types of sensors, including but not limited to SERS.
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A facile synthetic strategy is devised to construct raspberry like gold nanoparticles (RbNPs) formed by gold nanoclusters wrapped around ß-cyclodextrin functionalized gold nanoparticles (CD-AuNPs@AuNCs). An efficient and sensitive electrochemical sensor for the detection of Cr(VI) has been developed based on RbNPs. The sensing platform exhibits an excellent wide linear range (100â pg mL-1 to 10â µg mL-1 ), extremely low detection limit (40.91â fg mL-1 i. e. 0.79â pM), which may pave a new way to fabricate other ultrasensitive electrochemical sensors based on the designed RbNPs.
Assuntos
Ciclodextrinas , Nanopartículas Metálicas , Rubus , Ouro , ÍonsRESUMO
BACKGROUND: This work was designed to explore the roles of PIM-1 in the development of cervical cancer. METHODS: There were 90 paired cervical tumor samples and the non-tumor adjacent tissue. The levels of PIM-1 in different samples were examined using quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) methods. The potential diagnostic value of PIM-1 was analyzed by the receiver operating characteristic (ROC) curve; furthermore, the expression of EGFR in tumor samples was detected, and Pearson's correlation analysis was performed to analyze the relationship between the expression of PIM-1 and EGFR. Finally, cervical cancer cell line Hela cells were cultured and treated by PIM-1 siRNA, and MTT assay and Pi/Annexin V assay were performed to explore the effects of PIM-1 siRNA on the growth and apoptosis ability of the Hela cells. RESULTS: PIM-1 was significantly up-regulated in cervical cancer tissue compared to adjacent tissue, and the expression of PIM-1 in patients with cervical cancer is positively associated with the size and metastasis of the tumor. ROC analysis showed PIM-1 is a sensitive biomarker for the diagnosis of cervical cancer. Furthermore, EGFR was over-expressed in cervical cancer tumor tissues, and the levels of PIM-1 and EGFR in cervical cancer tissue were positively correlated. Finally, PIM-1 siRNA dramatically inhibited the viability and promoted the apoptosis of the Hela cells. CONCLUSION: Our findings prove that PIM-1 may function as an oncogene in cervical cancer and can regulate the EGFR signaling in cervical cancer.
Assuntos
Oncogenes/genética , Proteínas Proto-Oncogênicas c-pim-1/metabolismo , Neoplasias do Colo do Útero/genética , Receptores ErbB/metabolismo , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Transdução de Sinais , TransfecçãoRESUMO
Selective profiling of steviol-catalyzing UDP-glycosyltransferases in plants was accomplished with a probe metabolically synthesized from two substrate-derived components comprising an alkynylated sugar receptor (steviol) module and a diazirine-modified sugar donor (UDP-glucose) module, thereby illustrating a facile approach for harnessing biosynthetic enzymes of natural glycosides in plants for synthetic biology.
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Diterpenos do Tipo Caurano/metabolismo , Glucuronosiltransferase/metabolismo , Sondas Moleculares/metabolismo , Alcinos/química , Arabidopsis/enzimologia , Proteínas de Arabidopsis/metabolismo , Produtos Biológicos/química , Produtos Biológicos/metabolismo , Diazometano/química , Diterpenos do Tipo Caurano/química , Sondas Moleculares/química , Especificidade por Substrato , Difosfato de Uridina/química , Difosfato de Uridina/metabolismoRESUMO
Cobalamin-independent methionine synthases (MS) are zinc-binding methyltransferases that catalyze de novo methionine biosynthesis in higher plants, which are enzymes critically involved in seed germination and plant growth. Here, we report a highly selective sulfonyl fluoride-based probe for chemoproteomic profiling of MS enzymes in living systems of the model plant Arabidopsis thaliana, as implemented in in-gel-, mass spectrometry-, and imaging-based platforms. This probe holds promise for facilitating and accelerating fundamental research and industrial application of MS enzymes, particularly in the contexts of MS1/2-targeting herbicide screening and design.
Assuntos
5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/química , Proteínas de Arabidopsis/química , Arabidopsis/enzimologia , Vitamina B 12/metabolismo , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/metabolismo , Sequência de Aminoácidos , Arabidopsis/química , Arabidopsis/genética , Arabidopsis/metabolismo , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Espectrometria de Massas , Alinhamento de Sequência , Ácidos Sulfínicos/química , Ácidos Sulfínicos/metabolismo , Vitamina B 12/químicaRESUMO
Cancer cells rely on fatty acid synthase (FASN), a key enzyme for de novo biosynthesis of long chain fatty acids, to sustain their proliferative potential and drive invasion. Unfortunately, conventional FASN assays are technically inadequate for discerning otherwise elusive FASN activity in complex biological milieux, which has hindered progress in the functional study of FASN and development of its inhibitors. Here, we describe a chemical probe with unprecedented selectivity and sensitivity for the labeling of active FASN in living cells, thus demonstrating a new analytical modality for visualizing endogenous FASN activity and exploring opportunities for drug discovery.