64 BIOTECHNOLOGY
Fig. 2. Interaction between antigen A and shMFE conjugated NPs and free shMFE.
As evidenced from the binding curves – shown in Fig. 1. (a) and (b) – the surface charge of the NPs clearly influence the binding kinetics. Te results show the applicability of Attana’s innovative cell-based biosensor system for the studies of interactions at the bio-nano interface.
Furthermore, the results indicate that this platform could also be used to investigate the impact of the physiochemical properties of NPs on their bio-molecular interactions.
Determination of kinetics and affinity of NPs interaction Due to the ambiguity of molecular weight of the corona coated particles, the association rate and affinity can only be calculated after weight determination of the particles.
Alternatively, the kinetic parameters and affinity of the interactions between NPs and their receptors can be calculated if the system is reversed and set in a biochemical assay.
Experiments have been conducted using the standard biochemical format, where one binding partners is immobilised onto the sensor surface.
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Fig. 2. shows the interaction between antigen A and shMFE conjugated NPs and free shMFE.
shMFE conjugated NP was immobilised onto the sensor surface using amine coupling chemistry and binding was studied by injecting the antigen A at five concentrations in duplicates.
After each sample injection the sensor chip was regenerated. Binding of free shMFE onto antigen A was also investigated following the above procedure. Raw data was processed using evaluation software, a part of the Attaché software suite. Data used for deriving kinetic rate constants was referenced by subtracting data from buffer injections and referenced with channel B. Kinetic rate constants were derived using a 1:1 binding model.
Te derived kinetic rate constants and the calculated affinity of the two interactions are very similar, indicating the recognition of shMEF by antigen A. Te process of conjugation does not seem to interfere with the interaction of
the two molecules, see Fig. 2. (a), (b). As evidenced from Fig. 2., the kinetic rate constants and affinity values are associated with small errors.
What can we learn from these results? In conclusion, the cell-based and biochemical assay platforms Attana has developed have demonstrated the potential to become the technique of choice when investigating the interactions at bio-nano interfaces. Tis approach can be used in more detailed experiments concerning bio- nano interface, for instance identification of binding partners of the NPs.
Te label-free Attana Cell 200 biosensor is a key component of Attana’s vision to provide the ability to measure molecular kinetic interactions in biologically relevant systems. Te ‘one-stop-shop’ instrument makes it possible to combine the strengths of both high quality biochemical and cell-based assays in one system.
For more information ✔ at
www.scientistlive.com/eurolab
Teodor Aastrup, Diluka Peiris & Daniel Wallinder are with Attana in Stockholm, Sweden.
www.attana.com
REFERENCES: 1
Aastrup, T (2013).
Innovations in Pharmaceutical Louise L., Käck, C., Aastrup,
Technology, pp. 48-51 2
T., Nicholls, I.A (2015). Sensors. Peiris, D., Markiv, A.,
15, 5884-5894 3
Curley, P., Dwek, M.V. (2012) Biosensors and Bioelectronics. 35:160-166
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