Question

In: Biology

we are planning to design a fluorescent-based biosensor to detect Ochratoxin A in dry wheat samples,...

we are planning to design a fluorescent-based biosensor to detect Ochratoxin A in dry wheat samples, our recognition element is Biotinylated aptamer. what is the immobilization method used in this case? how to describe it?

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Expert Solution

Ans. Aptamers are the small single stranded sequences of DNA / RNA being used to specifically bind the analytes to detect the analytes/ target in the sample. Thus aptamers can be used as recognition element/ detecting agent/ receptor in the biosensors/ analytic devices. For this, aptamers need to be fixed/ immobilized on a surface.

Immobilization of aptamer can be done by several methods mainly by covalent cross linking or by adsorption of aptamer on the substrate/ surface or modified/ treated surface.

1. Attachment od aptamer to gold particle/ film with help of linkers or on the modified surface like silica/ silica oxide by covalent bonding.

2. Biotin labelled aptamers can be attached to avidin/ steptavidin/ neutravidin protein coated surfaces like magnetic beads/ gold nanoparticles/ gold films etc  based on the type to transducer/ detector being used. Because biotin strongly binds non-covalently with strepavidin, thats why it can be used to immobilize the biotinylated aptamers. The dissociation constant for biotin (vitamin7 or vitamin H ) with streptavidin is around 10-15 mol/L.

Thus immobilization of biotinylated aptamer on the strepavidin coated surfaces/ columns/ naniparticles/ films is the better option.


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