How long you immobilized it? when its interact with the crosslinking agent, it needs more time (I did 15 hours at 4 C to immobilize cholesterol oxidase enzyme on magnetite and the yield could achieve higher than 60%).

Meka Saima

Thanks Meka. How about incubation time with glutaraldehyde? How long did it take and in what temperature?

Dear Fateme Mamashli,

This is the major problem who uses to work in enzyme immobilization project. But do not worry, Immobilization of Horseradish Peroxidase on Fe3O4 nanoparticle can be enhanced by performing the batch optimization studies in order to demonstrate the effects of initial solution pH, GA concentration, APTS concentration, temperature, and time of the immobilization process on the relative activity of the immobilized enzyme

Also, in Enzyme-immobilized magnetic nanoparticles study, the synthesis of Fe3O4 nanoparticle need to be optimised in order to enhance the unique properties such as large surface area, high bioactivity, and excellent stability which play a very important role in Immobilization of Horseradish Peroxidase.

Best of Luck

Fateme Mamashli The incubation time was only for 2 h at 25 C.

Hope you can get good result.

and I do agree with Sadanand Pandey , the synthesis method need to be improved to get unique properties. But it may also could decreasing the activity.

I was trying to get the materials back after immobilization process, and use it for another case.