Browsing by Author "Minunni, Maria E."
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Item Open Access Approaches to allergy detection using aptasensors(Taylor and Francis, 2007) Brys, Emile; Tombelli, Sara; Minunni, Maria E.; Mascini, Marco; Turner, Anthony P. F.1. Immunoglobulins and IgE Antibodies are glycoprotein molecules which are produced by plasma cells in response to an immunogen (1). They belong to a class of spherical proteins called globulins and are hence known as immunoglobulins (Igs). Their primary function is to mediate the host immune response by binding to antigens.Item Open Access Detection of TP53 mutation using a portable Surface Plasmon Resonance DNA-based biosensor(Elsevier, 2005-02-02T17:36:22Z) Jiang, Tieshan; Minunni, Maria E.; Wilson, P. K.; Zhang, Jian; Turner, Anthony P. F.; Mascini, MarcoA DNA-based Surface Plasmon Resonance (SPR) biosensor has been developed for the detection of TP53 mutation using the inexpensive and commercially available instrument, SPREETATM SPR-EVM-BT, from Texas Instruments. A direct immobilization procedure, based on the coupling of thiol-derivatised oligonucleotide probes (Probe-C6-SH) to bare gold sensor surfaces, was optimized using synthetic oligonucleotides. Hybridization reactions between the immobilized probe and a short sequence (26 mer) complementary, non-complementary and one point mutation DNA were then investigated. The main analytical parameters of the sensor system were studied in detail including selectivity, sensitivity, reproducibility and analysis time. Finally, the sensor system was successfully applied to polymerase chain reaction (PCR) amplified real samples, DNA extracted from both normal, wild type, (Jurkat) and mutated (Molt 4), carrying the mutation at codon 248 of the TP53 cell lines. The results obtained demonstrate that the DNA-based SPR biosensor was able to distinguish sequences present in the various samples that differ only by one base and hence it appears to be a strong candidate technique for the detection of gene mutation.Item Open Access A novel optical biosensor format for the detection of clinically relevant TP53 mutations(Elsevier, 2005-05-15) Wilson, P. K.; Jiang, T.; Minunni, Maria E.; Turner, Anthony P. F.; Mascini, MarcoThe TP53 gene has been the subject of intense research since the realisation that inactivation of this gene is common to most cancer types. Numerous publications have linked TP53 mutations in general or at specific locations to patient prognosis and therapy response. The findings of many studies using general approaches such as immunohistochemistry or sequencing are contradictory. However, the detection of specific mutations, especially those occurring in the structurally important L2 and L3 zinc binding domains, which are the most common sites of TP53 mutations, have been linked to patient prognosis and more strongly to radiotherapy and chemotherapy resistance in several major cancers. In this study, the TI-SPR-1 surface plasmon resonance system and Texas Instruments Spreeta™ chips were used to develop a DNA biosensor based on thiolated probes complementary to these domains. The sensors were able to detect these mutations in both oligonucleotides and PCR products with normal and mutant TP53 DNA, but the difference in hybridisation signal was small. Preliminary experiments to enhance the signal using Escherichia coli mismatch repair proteins, MutS and single strand binding protein were carried out. It was found that MutS was unable to bind to mismatch oligonucleotides, but single strand binding protein was able to bind to single stranded probes, which had not hybridised to the target, resulting in a three-fold increase in the sensitivity of the biosensor. While further work needs to be carried out to optimise the system, these preliminary experiments indicate that the TI-SPR-1 can be used for the detection of clinically relevant mutations in the TP53 gene and that the sensitivity can be increased significantly using single strand binding protein. This system has a number of advantages over current mutation detection technologies, including lower cost, ease of sensor preparation and measurement procedures, technical simplicity and increased speed due to the lack of need for gel electrophoresis.Item Open Access Surface plasmon resonance imaging for affinity-based biosensors(Elsevier, 2010-01-15) Scarano, Simona; Mascini, Marco; Turner, Anthony P. F.; Minunni, Maria E.SPR imaging (SPRi) is at the forefront of optical label-free and real-time detection. It offers the possibility of monitoring hundreds of biological interactions simultaneously and from the binding profiles, allows the estimation of the kinetic parameters of the interactions between the immobilised probes and the ligands in solution. We review the current state of development of SPRi technology and its application including commercially available SPRi instruments. Attention is also given to surface chemistries for biochip functionalisation and suitable approaches to improve sensitivity.