Parameters necessary to define an immobilized enzyme preparation

[Display omitted] •Understanding of the enzyme immobilization is required to optimize the processes.•Immobilization parameters and courses become critical to understand the process.•Changes in enzyme properties during immobilization can complicate the calculations.•Likely artifacts and problems to make these calculations are discussed.•Some advices to improve the understanding of immobilization processes are given. Biocatalytic processes continue to find increasing application in industry. Therefore enzyme immobilization has also become of increasing importance as a means of allowing enzyme containment within reactors operating in continuous mode or else separation of enzyme after use in (fed-)batch reactors, as well as potential recycle. Whilst much has been reported in the scientific literature about enzyme immobilization methods, in many cases the protocol leads to losses in enzyme activity. In this review we outline the reasons for loss of activity during immobilization and highlight suitable diagnostic tests to elucidate the precise cause and thereby methods to restore activity. The need for standardized reporting of immobilization methods is also emphasized as a means of benchmarking alternative approaches.