The question is bit confusing. In fact, immobilization is a modern technique concerning with Enzymology is just to separate out the enzyme of interest from the mixture or an industrial effluent. For awareness, the two major types of 'Immobilization' of enzymes are: Immobilization On and Immobilization In.

Further sub-categorization reflects as: 'Adsorbtion' and 'Encapsulation'. The most effective is encapsulation that is widely being used.

For comprehensive technical account, an individual should go through certain research papers on "Immobilization of Enzymes".

In general, one step purification of an enzyme from the crude extract is affinity/ligand binding chromatography.But it is not applicable to enzyme that is already immobilized.

If the enzyme was immobilized on magnetic beads, it may be easier.

I do agree with the statement from Liangliang.

Thanks for your replies Sanjay Mishra and Liangliang. For example my enzyme is microencapsulated, i used this encapsulated enzyme for treating cotton fabrics (biopolishing), the effluent contains the encapsulated enzyme and this enzyme can be used for 5-6 cycles, how can i separate the enzyme and reuse it?

What are techniques used for separation of encapsulated enzyme from effluent?

In fact, this is not a theoretical question to answer for. The practical approach in this reference may be ligand-binding/affinity chromatographic technique as I mentioned earlier. Initially it may be hit and trial method, and after optimization it would certainly be a validated technique. It is my opinion based on experience gathered so far. This statement is open for others to discuss on for further clarification. I always welcome suggestions/comments on my scientific statement as it should be a collaborative approach to solve any scientific problem with positivity.

Immobilization on "Magnetic nanoparticles". One approach as example for similar immobilization is attached:

Article Magnetic Gold Nanoparticles in SERS-Based Sandwich Immunoass...

@ Arunas Ramanavicius: Thanks. Your suggestion would certainly be providing new insights for exploring our studies in the area of 'Enzymology'.