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?

Solutions

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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