Preparation of pepsin immobilized cryogel bioreactors

Cryogels are three-dimensional polymeric skeletal structures, bonded to each other by porous networks, permitting transport by convertion, and have been developed as new separation matrices to be applied in various bioseparation processes. In recent years, cryogels have also been used as bioreactors for the production and degradation of therapeutic proteins. Bioreactors are available in the health sector for diagnostic, therapeutic purposes and for single use in biopharmaceutical production. However, the single use of bioreactors significantly increases the cost and reduces reproducibility. In biopharmaceutical applications, it is a targeted approach to use the desired degradation products of proteins instead of all proteins. Thus, contamination from nonspecific interactions will be avoided. F (ab´) 2 fragments are the region of the immunoglobulin G (IgG) molecule used in antigen-antibody interactions. Commercially sold therapeutic monoclonal F (ab´) 2 fragments are very important in the health sector. In the present study, it has been studied in a continuous system and at the same time it has been suggested that an easy and effective cryogel bioreactor is made to immobilize the pepsin enzyme. For efficient and reliable design of Pepsin immobilization using cryogel bioreactor, covalent immobilization was performed between the epoxy groups of the matrix and the amino groups of Pepsin enzyme. The enzyme was characterized by immobilized cryogel bioreactor SEM and swelling test. The specific activity, substrate concentration, temperature and storage stability of the immobilized peps over a wide pH range were compared with the free pepsin in a solution using IgG susbstrate.