An enzymatic nucleic acid vertical flow assay.

Vertical flow assays have been developed in recent years addressing limitations of the lateral flow assays, including limited multiplexing capability, quantitation, and hook effect. In the present study, the first passive paper-based vertical flow assay is developed for the detection of the nucleic acid target. Horseradish peroxidase was used as an enzymatic tracer with a high potential for signal amplification. In order to achieve the best signal-to-noise ratio, different parameters of paper-based assays were optimized. The sample is heat denatured and hybridized with a specific probe to form a dual-labeled hybridization complex. A small volume of diluted sample, 12 µl, can be analyzed within 6 min on the assay in a sandwich format. Assay specificity was evaluated by testing different unrelated samples, and also, 1.7 nM was obtained as the limit of detection (LOD) using the 0 + 3SD method, which is equivalent to 8.5 fmol of double-stranded DNA in the 12 µl sample volume. The linear range of 3-194 nM with a 0.978 correlation coefficient was obtained according to the calibration curve. The developed assay was evaluated with 45 hepatitis B virus clinical plasma samples, and the result showed 100% consistency of the assay with the real-time PCR benchmark. In the present study, we sought to develop a mere detection system for nucleic acid targets, and to investigate the possibility of using enzyme reporter in a passive vertical flow assay.

Angiogenic biomolecules specific nanobodies application in cancer imaging and therapy; review and updates.

Solid cancers are affiliate on angiogenesis for preservation. The FDA-approved monoclonal antibodies like Bevacizumab are currently being administered effectively as inhibitors of angiogenesis against various types of tumors. Despite the clinical achievements in this regard, monoclonal antibodies suffer from considerable disadvantages, including the potential to develop therapeutic resistance, a high production cost, and the restricted tumor penetration, which consequently limit their therapeutic applications. In the past decades, some significant methods such as miniaturization of the antibodies, containing those inhibiting tumor angiogenesis, have been proposed to enhance cancer therapeutics efficiency. Since their discovery, small single-domain antigen binding antibody fragments, known as nanobodies, have been broadly utilized in the fields of cancer research, diagnosis, and treatment. Due to their desired functional attributes and a unique structure, nanobodies are becoming promising therapeutic and diagnostic biomolecules in oncology field. Moreover, they displayed a substantial translational potential in preclinical and clinical studies. This review was performed with the aim of highlighting the potential of nanobodies in blocking the angiogenic process by targeting of angiogenic biomolecules for cancer therapy and the application of labeled nanobodies in non-invasive in vivo tumor imaging.

Delivery of Various Cargos into Cancer Cells and Tissues via Cell-Penetrating Peptides: A Review of the Last Decade.

Cell-penetrating peptides (CPPs), also known as protein transduction domains, are a class of diverse amino acid sequences with the ability to cross cellular membranes. CPPs can deliver several bioactive cargos, including proteins, peptides, nucleic acids and chemotherapeutics, into cells. Ever since their discovery, synthetic and natural CPPs have been utilized in therapeutics delivery, gene editing and cell imaging in fundamental research and clinical experiments. Over the years, CPPs have gained significant attention due to their low cytotoxicity and high transduction efficacy. In the last decade, multiple investigations demonstrated the potential of CPPs as carriers for the delivery of therapeutics to treat various types of cancer. Besides their remarkable efficacy owing to fast and efficient delivery, a crucial benefit of CPP-based cancer treatments is delivering anticancer agents selectively, rather than mediating toxicities toward normal tissues. To obtain a higher therapeutic index and to improve cell and tissue selectivity, CPP-cargo constructions can also be complexed with other agents such as nanocarriers and liposomes to obtain encouraging outcomes. This review summarizes various types of CPPs conjugated to anticancer cargos. Furthermore, we present a brief history of CPP utilization as delivery systems for anticancer agents in the last decade and evaluate several reports on the applications of CPPs in basic research and preclinical studies.