Immobilized nanoparticles-mediated enzyme therapy; promising way into clinical development.

Enzyme (Enz)-mediated therapy indicated a remarkable effect in the treatment of many human cancers and diseases with an insight into clinical phases. Because of insufficient immobilization (Imb) approach and ineffective carrier, Enz therapeutic exhibits low biological efficacy and bio-physicochemical stability. Although efforts have been made to remove the limitations mentioned in clinical trials, efficient Imb-destabilization and modification of nanoparticles (NPs) remain challenging. NP internalization through insufficient membrane permeability, precise endosomal escape, and endonuclease protection following release are the primary development approaches. In recent years, innovative manipulation of the material for Enz immobilization (EI) fabrication and NP preparation has enabled nanomaterial platforms to improve Enz therapeutic outcomes and provide low-diverse clinical applications. In this review article, we examine recent advances in EI approaches and emerging views and explore the impact of Enz-mediated NPs on clinical therapeutic outcomes with at least diverse effects.

SARS-CoV-2 mRNA-vaccine candidate; COReNAPCIN®, induces robust humoral and cellular immunity in mice and non-human primates.

At the forefront of biopharmaceutical industry, the messenger RNA (mRNA) technology offers a flexible and scalable platform to address the urgent need for world-wide immunization in pandemic situations. This strategic powerful platform has recently been used to immunize millions of people proving both of safety and highest level of clinical efficacy against infection with severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Here we provide preclinical report of COReNAPCIN®; a vaccine candidate against SARS-CoV-2 infection. COReNAPCIN® is a nucleoside modified mRNA-based vaccine formulated in lipid nanoparticles (LNPs) for encoding the full-length prefusion stabilized SARS-CoV-2 spike glycoprotein on the cell surface. Vaccination of C57BL/6 and BALB/c mice and rhesus macaque with COReNAPCIN® induced strong humoral responses with high titers of virus-binding and neutralizing antibodies. Upon vaccination, a robust SARS-CoV-2 specific cellular immunity was also observed in both mice and non-human primate models. Additionally, vaccination protected rhesus macaques from symptomatic SARS-CoV-2 infection and pathological damage to the lung upon challenging the animals with high viral loads of up to 2 × 108 live viral particles. Overall, our data provide supporting evidence for COReNAPCIN® as a potent vaccine candidate against SARS-CoV-2 infection for clinical studies.

A Comparative Analysis of Different Enzyme Immobilization Nanomaterials: Progress, Constraints and Recent Trends.

Immobilization techniques have been popularly used to preserve the operational stability of the enzymes for industrial applications. The three main components of an immobilized enzyme system are the enzyme, the matrix/support, and the technique of immobilization. So far, different supports have been developed to improve the efficiency of the immobilized enzymes. But in the recent decade, nanotechnology has been of considerable research interest in the field of immobilized enzyme carriers. The materials at the nano-scale, due to their unique physicochemical properties, including; specific surface area, mass transfer limitation, and effective enzyme loading, are considered interesting matrices for enzyme immobilization. This review describes techniques employed to immobilize enzymes, and provides an integrated focus on the most common nanoparticles for enzyme conjugation. Additionally, the pros and cons of nanoparticles as immobilization matrices are also discussed. Depending on the type of enzyme and its application, in this review, the researchers are directed to select an appropriate method and support for enzyme immobilization in terms of enzyme stability and functionality.

Immobilization of lactoperoxidase on graphene oxide nanosheets with improved activity and stability.

OBJECTIVES: The purpose of this study was to develop a facile and efficient method to enhance the stability and activity of lactoperoxidase (LPO) by using its immobilization on graphene oxide nanosheets (GO-NS). METHODS: Following the LPO purification from bovine whey, it was immobilized onto functionalized GO-NS using glutaraldehyde as cross-linker. Kinetic properties and stability of free and immobilized LPO were investigated. RESULTS: LPO was purified 59.13 fold with a specific activity of 5.78 U/mg protein. The successful immobilization of LPO on functionalized GO-NS was confirmed by using dynamic light scattering (DLS) and Fourier transform infrared spectroscopy (FT-IR). The overall results showed that the stability of the immobilized LPO was considerably improved compared to free LPO. Apparent Km and Vmax of LPO also indicated that the immobilized enzyme had greater affinity to the substrate than the native enzyme. CONCLUSIONS: Graphene oxide nanosheets are effective means for immobilization of LPO.

Purification of lactoperoxidase from bovine whey and investigation of kinetic parameters.

BACKGROUND: Lactoperoxidase (LPO) is related to mammalian peroxidase family which contains a wide spectrum of biological activities. Despite the wide studies on the LPO, there is little study has been performed to simplify and shorten the procedure of enzyme purification. The aim of this project was to purify the enzyme through a simple method, and investigating enzyme kinetic parameters. MATERIALS AND METHODS: LPO was purified from bovine whey through modified method of Yoshida (1990) using two steps of ammonium sulfate precipitation and ion-exchange chromatography. The purity of isolated enzyme was monitored by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). RESULTS: The enzyme was purified 59.13-fold with a recovery of 10.26 having a specific activity of 5.78 U/mg protein and an Rz value of 0.8. The enzyme activity was measured using guaiacol as a chromogenic substrate in phosphate buffer pH 6. SDS-PAGE showed a single bond with molecular weight of 78 kDa. The purified enzyme displayed optimum activity at pH 6 in 30 mM phosphate buffer and at a temperature of 50°C, with a Km value of 178 mM and Vmax 0.63 U/ml.min for guaiacol. CONCLUSION: Using only one step ion-exchange chromatography, LPO was isolated from bovine whey in high purity.