NOVEL SMART pH SENSITIVE CHITOSAN GRAFTED ALGINATE HYDROGEL MICROCAPSULES FOR ORAL PROTEIN DELIVERY: II. EVALUATION OF THE SWELLING BEHAVIOR

Objective: The main objective of this study is to evaluate the swelling behavior of pH sensitive chitosan (CS) grafted alginate (ALG) hydrogel microcapsules and compared with a simple alginate-chitosan mixed polyelectrolyte complex (PEC) to show the benefits of the used covalently grafting technique. In addition, the behavior of the swelling process under physiological conditions to stimulate gastric, colonic and intestinal medium for grafted PEC microcapsules will be investigated as well.Methods: The new pH sensitive hydrogel microcapsules were prepared using grafting to†technique. Swelling studies were conducted in buffer saline solutions with different pHs using wet beads. In addition; the sensitivity of the grafted microcapsules to the change of pH in the simulated gastric fluid (SGF; pH 1.2), (SIF; pH 6.8) and (SCF; pH 7.4) was investigated.Results: It was observed from the swelling studies that sharp phase transition was recognized between pH 3–4. While this transition became broader and recognized between pH 3.0-7.4, where the maximum value of the equilibrium swelling degree was varied depending on the variation of CS concentration from 0.1% to 0.5%, both grafted and mixed microcapsules exhibit higher swelling degree at high pH 6.8 (120%, 100%) respectively.Conclusion: It was clear from all swelling studies that the grafting technique may be a suitable way for large-scale production of pH sensitive alginate–chitosan microcapsules as a potential system for site-specific oral delivery of protein drugs to different regions of the intestinal tract.Â

NOVEL SMART pH SENSITIVE CHITOSAN GRAFTED ALGINATE HYDROGEL MICROCAPSULES FOR ORAL PROTEIN DELIVERY: I. PREPARATION AND CHARACTERIZATION

Objectives: Preparation and characterization of a new pH sensitive chitosan (CS) grafted alginate (ALG) hydrogel microcapsules for the oral delivery of protein.Methods: The pH sensitive hydrogel microcapsules were prepared for the first time using grafting to†technique. Firstly, alginate was activated using Ï-Benzoquinone (PBQ) as a coupling agent to graft Chitosan chains later on. Both of activated and grafted alginate microcapsules were characterized by Fourier transform-Infra red spectroscopy (FT-IR), thermal gravimetric analysis (TGA) and the morphological structures were investigated using Scanning electron microscopy (SEM) examination.Results: It was found that the optimum conditions affecting the activation process and also the swelling degree of the prepared hydrogel microcapsules were 2% ALG, 0.04M PBQ pH10, 45 °C for 2h. In addition, the grafting process depends on the attached amount of PBQ and CS concentration. Maximum grafting efficiency (GE %) and chitosan add-on percentage were 98.6% and 14.8% respectively using 0.3% CS at 40 °C for 3h.Conclusions: Novel pH sensitive hydrogel microcapsules were prepared via grafting of chitosan molecules on to activated alginate backbone. The formulated microcapsules can be applied as a new pH sensitive carrier for protein drugs. Â

Hydrogel coated monoliths for enzymatic hydrolysis of penicillin G

The objective of this work was to develop a hydrogel-coated monolith for the entrapment of penicillin G acylase (E. coli, PGA). After screening of different hydrogels, chitosan was chosen as the carrier material for the preparation of monolithic biocatalysts. This protocol leads to active immobilized biocatalysts for the enzymatic hydrolysis of penicillin G (PenG). The monolithic biocatalyst was tested in a monolith loop reactor (MLR) and compared with conventional reactor systems using free PGA, and a commercially available immobilized PGA. The optimal immobilization protocol was found to be 5 g l−1 PGA, 1% chitosan, 1.1% glutaraldehyde and pH 7. Final PGA loading on glass plates was 29 mg ml−1 gel. For 400 cpsi monoliths, the final PGA loading on functionalized monoliths was 36 mg ml−1 gel. The observed volumetric reaction rate in the MLR was 0.79 mol s−1 m−3monolith. Apart from an initial drop in activity due to wash out of PGA at higher ionic strength, no decrease in activity was observed after five subsequent activity test runs. The storage stability of the biocatalysts is at least a month without loss of activity. Although the monolithic biocatalyst as used in the MLR is still outperformed by the current industrial catalyst (immobilized preparation of PGA, 4.5 mol s−1 m−3catalyst), the rate per gel volume is slightly higher for monolithic catalysts. Good activity and improved mechanical strength make the monolithic bioreactor an interesting alternative that deserves further investigation for this application. Although moderate internal diffusion limitations have been observed inside the gel beads and in the gel layer on the monolith channel, this is not the main reason for the large differences in reactor performance that were observed. The pH drop over the reactor as a result of the chosen method for pH control results in a decreased performance of both the MLR and the packed bed reactor compared to the batch system. A different reactor configuration including an optimal pH profile is required to increase the reactor performance. The monolithic stirrer reactor would be an interesting alternative to improve the performance of the monolith-PGA combination

Immobilization of enzymes on supports activated with glutaraldehyde: A very simple immobilization protocol

In this chapter, we describe different approaches for the utilization of glutaraldehyde in protein immobilization. First, we focus on the covalent attachment of proteins to glutaraldehyde-activated matrixes. We describe conditions for the synthesis of such supports and provide an example of the immobilization and stabilization of a fructosyltransferase. We also describe how glutaraldehyde may be used for the cross-linking of protein–protein aggregates and protein adsorbed onto amino-activated matrixes. In these cases, glutaraldehyde bridges either two lysine groups from different protein molecules or a lysine from the protein structure and an amine group from the support. Examples of cross-linking are given for the immobilization of a D-amino acid oxidase on different amino-activated supports.Peer reviewe