What are the limitations of covalent binding immobilization?

Covalent binding is an enzyme immobilization technique that involves connecting enzymes with support materials such as agarose, porous silica, or polyacrylamide, via strong, highly stable linkages. 

Disadvantages of Covalent Binding:

• Chemical alteration of the enzyme can lead to loss in functional conformation
• Enzymes get inactivated due to conformation changes when active site reactions occur