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Molecularly Imprinted Polymers in Biotechnology
Very few of the tens of reviews already published on this subject present a critical analysis of the technological aspects of molecular imprinting. Leaders in this field have been approached with requests to provide their views and analyses of specific areas of design, characterization and application of these polymers. The last, but by no means least, part of the book is dedicated to often overlooked associated aspects of MIPs such as commercialization strategy and IPR, prepared by Peter Leverkus and Jeffrey McIntyre.
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Fulfilment Centre Email: siamparagon kinokuniya. Check Retail Stores' phone number. Data were graphically plotted and statistically analyzed using GraphPad Prism v7. Datasets for other assays were technical replicates within single experiments. Whilst infectious virus was detected in all sucrose gradient fractions, the highest titers were detected, as predicted, in the central fractions of the gradient, i.
Testing of the individual sucrose gradient fractions confirmed the highest titers in fractions 16—18 Figure 1B and these fractions were dialyzed into PBS and inactivated prior to imprinting. Infectious virus titers post-dialysis and post-inactivation were determined data not shown and electron microscopy confirmed the structural integrity of the template virus and the absence of significant cellular debris Figure 1C.
A PBS control treatment was used as a further negative control in each assay. There was no inter-batch variation in this effect suggesting a reproducible imprinting-related neutralization of virus. However, pNIPAM produced a very similar reduction in infectious viral titer in both its MIP and NIP forms, suggesting that molecular imprinting was not entirely responsible for the neutralization event in the case of this polymer. Figure 1.
Template virus was prepared by purification of PRRSV-1 virions by continuous density ultracentrifugation. Pools of gradient fractions A and individual fractions B containing the highest infectious titers were selected and virion purity and integrity post-dialysis and inactivation confirmed by electron microscopy C. The limit-of-detection of the assays are indicated by dashed horizontal lines.
Both MIPs showed a broadly similar effect with decreasing concentration, providing complete neutralization of infectious virus to the LoD of the assay at both neat and dilutions. Figure 2. Characterization of the virus neutralizing properties of hydrogel MIPs. The limit-of-detection of the assays are indicated by horizontal dashed lines. This is the first study to demonstrate that MIPs imprinted with a clinically relevant virus can exert potent antiviral effects, reducing the infectious viral titer recovered to below the LoD of the assay used.
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The concentration and time dependent effect of neutralization further supports a rapid and specific binding. That there is a virus neutralization event in this study, regardless of imprinting taking place, points to the potential toxic nature of pNIPAM. It has been established in this study with an animal virus and by others with bacteriophages Sankarakumar and Tong, ; Li et al.
Molecularly Imprinted Polymers in Biotechnology by Bo Mattiasson, Paperback | Barnes & Noble®
The results of our incubation time trial showing that complete neutralization could be achieved in as little as 2. However, this would need to be re-assessed in the context of plasma proteins and other potentially interfering molecules that would be present in vivo. In terms of suitability for in vivo testing, further work is needed to ensure suitable biocompatibility of our MIPs. Li et al. The latter demonstrated the ability of MIPs to bind the cytotoxic peptide melittin, the principle component of bee venom, in the bloodstream of mice, which significantly reduced the mortality and morbidity associated with melittin envenomation.
The data from this study has demonstrated a highly effective and specific neutralization of virus infectivity with certain hydrogel-based MIPs. Whilst promising, it is possible that the destructive method used to produce these cavity-containing hydrogel-MIPs leads to the majority of the material comprising redundant unselective particles, devoid of template-specific cavities.
Further studies will evaluate virus imprinting of nanoparticle-based MIPs nanoMIPs for the efficient production of high bioaffinity materials Canfarotta et al. MIPs imprinted with virions may be produced according to a variety of methods, giving nanoscale shells with cavity populated surfaces. In conclusion, hydrogel-based MIPs are capable of specifically neutralizing virus infectivity in vitro within a short enough incubation time to be clinically relevant. All authors contributed to manuscript revision, read, and approved the submitted version.
The funders had no role in study design, data collection and interpretation, or the decision to submit the work for publication. The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.
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