Surface plasmon resonance biosensor assay for the analysis of small-molecule inhibitor binding to human and parasitic phosphodiesterases

Siderius, Marco; Shanmugham, Anitha; England, Paul; Meer, Tiffany van der; Bebelman, Jan Paul; Blaazer, Antoni R.; Esch, Iwan J. P. de; Leurs, Rob

doi:10.1016/j.ab.2016.03.013

Cited by 7 publications

(6 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…21 Understanding the interaction of LMW compounds is crucial in the design of new pharmaceuticals. 22 LMWs that absorb light in the wavelength used for SPR measurements allow a more sensitive detection. 21 Indeed, the ability of graphene to absorb light at visible wavelengths has motivated a series of theoretical 23 and experimental 12,24−28 investigations on using graphene on gold as a sensor surface when using SPR.…”

Section: Introductionmentioning

confidence: 99%

Binding Kinetics of Methylene Blue on Monolayer Graphene Investigated by Multiparameter Surface Plasmon Resonance

et al. 2018

View full text Add to dashboard Cite

In this paper, we study the interaction of a small dye molecule, namely, methylene blue (MB) with graphene surfaces using surface plasmon resonance (SPR). We show that by utilizing all of the parameters of the SPR angular dip and exploiting the fact that MB absorbs light at the operating wavelength, it is possible to detect the binding of small molecules that would otherwise not give a significant signal. The binding of MB to unmodified graphene is found to be stronger than that for gold. By studying the interaction at modified surfaces, we demonstrate that electrostatic effects play a dominant role in the binding of MB on to graphene. Furthermore, following the binding kinetics at various concentrations allows us to estimate apparent equilibrium binding and rate constants for the interaction of MB with graphene.

show abstract

Section: Introductionmentioning

confidence: 99%

Binding Kinetics of Methylene Blue on Monolayer Graphene Investigated by Multiparameter Surface Plasmon Resonance

et al. 2018

View full text Add to dashboard Cite

show abstract

“…The protein was stored in 50 mM Tris HCl pH 8.0, 100 mM NaCl, 2 mM DTT, 5 mM MgCl 2 and 5% glycerol.Phosphodiesterase activity. These assays were performed using the PDELight™ HTS cAMP phosphodiesterase Kit (Lonza, Walkersville, USA), at 25 °C in non-binding, low volume 384-well plates (Corning, Kennebunk, ME, USA), essentially as described[48]. SmPDE4A (0.6 mg/mL) activity measurements (cAMP concentration at 2xKm (6 µM)) were made in 'stimulation buffer' (50 mM HEPES, 100 mM NaCl, 10 mM MgCl 2 , 0.5 mM EDTA, 0.05 mg/mL BSA, pH 7.5).…”

mentioning

confidence: 99%

Discovery of Novel Schistosoma Mansoni PDE4A Inhibitors as Potential Agents against Schistosomiasis

et al. 2019

Self Cite

View full text Add to dashboard Cite

Aim: Due to the urgent need for effective drugs to treat schistosomiasis that act through a known molecular mechanism of action, we focused on a target-based approach with the aim to discover inhibitors of a cyclic nucleotide phosphodiesterase from Schistosoma mansoni (SmPDE4A). Materials & methods: To discover new inhibitors of SmPDE4A homology models of the enzyme structure were constructed based on known human and protozoan homologs. The best two models were selected for subsequent virtual screening of our in-house chemical library. Results & conclusion: A total of 25 library compounds were selected for experimental confirmation as SmPDE4A inhibitors and after dose-response experiments, three top hits were identified. The results presented validate the virtual screening approach to identify new inhibitors for clinically relevant phosphodiesterases.

show abstract

“…We also found that the bioactivity of human AFP protein is similar to that of cytoplasmic human AFP. Recombinantly expressed human AFP is soluble, is stable and can be used to identify signalling molecules or potential drug interactions by surface plasmon resonance (SPR) [24,25]. In conclusion, the present study demonstrated the successful expression and purification of R-AFP, which paves the way for future structural and functional studies of human AFP.…”

Section: Discussionmentioning

confidence: 57%

Expression and bioactivity of human α-fetoprotein in a Bac-to-Bac system

et al. 2017

View full text Add to dashboard Cite

α-fetoprotein (AFP) is an early serum growth factor in foetal embryonic development and hepatic oncogenesis. A growing number of investigations of AFP as a tumour-specific biomarker have concluded that AFP is an important target for cancer treatment. AFP also plays an immunomodulatory role in the treatment of several autoimmune diseases, such as rheumatoid arthritis, multiple sclerosis, myasthenia gravis and thyroiditis. In an effort to support biochemical screening and drug design and discovery, we attempted to express and purify human AFP in a Bac-to-Bac system. Two key factors affecting the expression of recombinant human AFP (R-AFP), namely the infectious baculovirus inoculum volume and the culturing time post-infection, were optimized to maximize the yield. We achieved a high yield of approximately 1.5 mg/l of harvested medium with a 72–96 h incubation period after infection and an inoculum volume ratio of 1:100. We also assessed the role of R-AFP in the proliferation of the human liver cancer cell line Bel 7402, and the results indicated that R-AFP promoted the growth of hepatoma cells. We concluded that this method can produce high yields of R-AFP, which can be used for studies related to AFP.

show abstract

Surface plasmon resonance biosensor assay for the analysis of small-molecule inhibitor binding to human and parasitic phosphodiesterases

Cited by 7 publications

References 34 publications

Binding Kinetics of Methylene Blue on Monolayer Graphene Investigated by Multiparameter Surface Plasmon Resonance

Binding Kinetics of Methylene Blue on Monolayer Graphene Investigated by Multiparameter Surface Plasmon Resonance

Discovery of Novel Schistosoma Mansoni PDE4A Inhibitors as Potential Agents against Schistosomiasis

Expression and bioactivity of human α-fetoprotein in a Bac-to-Bac system

Contact Info

Product

Resources

About