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Lab-on-a-Chip Immunoassay Systems
Published in Richard O’Kennedy, Caroline Murphy, Immunoassays, 2017
Barry Byrne, Louise M. Barrett
One of the key benefits of using microfluidics-based platforms is the ability to fabricate chips which permit parallel, multiplexed analysis, e.g. of bacterial cells and associated toxins. The latter include Staphylococcal enterotoxin B (SEB) [62] and cholera toxin [63], and due to the fact that they pose a considerable risk to human health, developing strategies for reliable and sensitive detection is also an important consideration. To demonstrate multiplexed analysis, the PDMS-based microfabricated immunoassay based on flow cytometry designed by Kim et al. [40], discussed earlier with reference to how the microfluidic design enhanced sample delivery, for the detection of bacterial cells (E. coli, Listeria spp. and Salmonella spp.) and toxins (SEB/cholera toxin and ricin). The sandwich assay format integrated fluorescently coded and carboxyfunctionalised microspheres, to which individual pathogen or toxin-specific antibodies were covalently coupled. Signal amplification was facilitated by the use of biotinylated, phycoerythrin-labelled anti-streptavidin antibodies, with optical fibres being integrated into the chip to facilitate laser-based detection. Limits of detection for bacterial cells were between 1×103 and 1×105 CFU mL−1. However, the ability to also integrate toxin-specific antibodies is a considerable advantage, which further demonstrates the potential of this and related platforms for multiplexed analysis of structurally diverse analytes. Toxin limits of detection for this study are summarised in Table 5.1, which also highlights other excellent examples of ‘lab-on-a-chip’ immunoassay-based bacterial toxin detection.
Design of artificial cells: artificial biochemical systems, their thermodynamics and kinetics properties
Published in Egyptian Journal of Basic and Applied Sciences, 2022
Adamu Yunusa Ugya, Lin Pohan, Qifeng Wang, Kamel Meguellati
Munshaw group work on a fully rational synthetic HCV subtype 1a virus (Bole1a) was designed by Bayesian phylogenetics, covariance analysis, and ancestral sequence reconstruction comprising mainly of 338 epitopes and envelope genes assembled and mediated in the entry into the target cells [60]. The intracellular incorporation of synthetic enhancers can help in the generation of viruses with novel properties and is also a good example cited in the construction of artificial cells [61]. Cottingham and his coworkers’ study is based on the clinically deployable viral vector by proof-of-concept experiments, modified Vaccinia virus Ankara bacterial artificial chromosomes (MVA-BCA), and it is found to be an effective new candidate for mutant and recombinant vaccines [62]. The construction of synthetic CMV promoters of all strengths was carried out by a 10-mer synthetic enhancer spacer [63]. A new approach for the treatment of DENV infection in humans is based on improving the nucleic acid inhibitors of the Dengue virus (DENV) by RNAi (RNA interference) based on multiple artificial microRNAs (amiRNAs) that target the conserved regions of the virus [64]. The few studies on artificial cells mimicking bacteria report the construction of Artesimisin artesunate (ARS), a synthetic tetraxane drug candidate (RKA182) and a trioxolane equivalent (FBEG100), which have been used as an efficient drug for the treatment of P. falciparum malaria [65]. Kissner and his group produced a synthetic dimeric BB-loop mimetic of MyD88 (EM-163) that inhibited pro-inflammatory signaling and was found to be toxic to Staphylococcal enterotoxin B therapeutically [66].