Nanomaterial-Supported Enzymes

The search for ideal solid support that promotes enzyme stability, easy separation and reuse cycles in different processes has grown widely. In this sense, core‐shell polymer particles draw attention because they are already widely used in adsorption, catalysis, and drug delivery, due to their remarkable advantages in surface properties adjustment, excellent mechanical stability, and high chemical resistance, as a result of the combination of the characteristics of the polymers that form the core and shell of the structure. Thus, this review article provides an overview of the synthesis processes of these particles, highlighting the characteristics that can be obtained from each synthesis technique, as well as the most recently used techniques to obtain polymeric particles with core‐shell morphology. The advantages and main challenges for the application of these structures in the immobilization of enzymes are also discussed, providing a general summary of what has been explored in the literature. Despite the potential of application, the use of these materials in the immobilization of enzymes is still little explored, with studies focused mainly on lipases and around the same classes of polymers. Therefore, there is an opportunity window regarding the investigation of the immobilization of other enzymes of commercial interest on these polymeric supports.

Enzymes play a fundamental role in the manufacture of biosensors, acting as biological recognition elements due to their sensitivity and selectivity in the reactions they catalyze. Among them, urease stands out as an efficient enzyme found in nature, whose function involves the hydrolysis of urea. This enzyme has been used when extracted from legume seeds, as well as various bacteria and fungi. The objective of this study was the extraction and immobilization of the urease enzyme in polyvinylpyrrolidone (PVP) films. Crude jack bean extracts were prepared at concentrations of 15, 25 and 35% in PBS/PVP, centrifuged and refrigerated. Subsequently, aliquots of each solution were applied to plates to form films, being characterized by enzymatic activity, pH, FTIR and UV-vis. The affinity with urea was evidenced by enzymatic activity, pH and UV-vis, compatible with the Michaelis-Menten model. In FTIR, the polypeptide bands characteristic of the amino acids present in the structure of the enzymes, such as those of amides I, II and III, were confirmed.

Nanotechnology is rapidly advancing and will leave no field untouched by its ground breaking innovations. Nanoparticles are molecules with a diameter ranging from 10-100 nm. Nanotechnology has promising biomedical applications and most noteworthy amongst them are noble metal particles. For instance, gold nanoparticles (AuNPs) provide a unique blend of physical and optical properties, chemical inertness, and high surface to volume ratio. They can be synthesized as well as functionalised to support various ligands on their surface. Their surface functionalization and diverse properties render the gold nanoparticles highly useful for drug delivery and gene carrier for therapeutic purposes and as molecular probes for disease diagnosis. The foundation for the usage of AuNPs in therapeutics and diagnosis was laid by the ancient studies done with ruby gold for curing diseases in middle ages. Presently, AuNPs have become available in different types such as spheres, rods, shells, cages and SERS particles which vary in shape, size and physical properties. The biomedical applications of these particles include drug and gene delivery, cancer diagnosis and therapy, determination of biological molecules and microorganisms, detection of disease etiology, immunoassay, enzyme immobilization, etc. Overall, the focus of this review is to highlight that AuNPs provide an excellent platform for the discovery of new therapies, cure for certain cancers, molecular probe for diagnostic purposes, as well as gene carriers and drug delivery vehicles. Biomed Rev 2015; 26: 23-36. Key words: gold nanoparticles, cancer treatment, drug delivery system, gold nanocarrier therapy

In a number of sectors, such as pharmaceuticals, food production, and environmental control, enzyme biotechnology is essential. Enzyme immobilization, which involves binding enzymes to solid supports, improves their stability, reusability, and efficiency. This review focuses on this procedure. The article examines the various techniques for immobilizing enzymes, including adsorption, covalent bonding, entrapment, and encapsulation. Each technique has special benefits, including improved process control, decreased contamination, and better stability. In pharmaceutical applications, immobilized enzymes are frequently utilized for high-purity active pharmaceutical ingredients (APIs), drug manufacturing, and enzyme-based drug delivery. The article also emphasizes their function in vaccine manufacture and biosensors for disease diagnostics. It also looks at how immobilized plant and bacterial cells can be used for environmental applications such as bioremediation, biotransformation, and the synthesis of secondary metabolites. Enzyme immobilization's overall importance stems from its capacity to maximize industrial processes, making them more economical, efficient, and sustainable. Enzymes continue to provide creative answers to a variety of biotechnological problems through a variety of immobilization approaches.

Bacterial cellulose (BC) is a versatile biopolymer with better material properties, such as purity, high degree of porosity, relative high permeability to liquid and gases, high water-uptake capacity, tensile strength and ultrafine network. This review explores the applications of BC and its hydrogels in the fields of food, cosmetics and drug delivery. Applications of BC in foods are ranging from traditional dessert, low cholesterol diet, vegetarian meat, and as food additive and dietary aid to novel applications, such as immobilization of enzymes and cells. Applications in cosmetics include facial mask, facial scrub, personal cleansing formulations and contact lenses. BC for controlled drug delivery, transdermal drug delivery, dental drug delivery, protein delivery, tissue engineering drug delivery, macromolecular prodrug delivery and molecularly imprinted polymer based enantioselective drug delivery are also discussed in this review. The applications of BC in food and cosmetics provide the basis for BC-based functional foods, nutraceuticals, cosmeceuticals and medicated cosmetics. On the basis of current studies, the BC-based drug delivery could be further fine-tuned to get more sophisticated control on stimuli-responsive drug release. Along with the currently available literature, further experiments are required to obtain a blueprint of drug in vivo performance, bioavailability and in vitro–in vivo correlation.

Current applications of Colloidal Liquid Aphrons: Predispersed solvent extraction, enzyme immobilization and drug delivery

The Covid-19 pandemic has brought about rapid change in medical science. The production of new generation vaccines for this disease has surprised even their most optimistic supporters. Not only have these vaccines proven to be effective, but the importance of this disease and pandemic situation also significantly shortened the long-standing process of validating such products. Vaccination is a type of immunotherapy. Researchers have long been looking at vaccines as a possible treatment for cancer (Geynisman et al., 2014). In the same way that vaccines work against infectious diseases, attempts are being made to develop vaccines to identify specific proteins on cancer cells. This helps the immune system recognize and attack cancer cells. Cancer vaccines may help: I) Prevent the growth of cancer cells (Bialkowski et al., 2016), II) Prevent recurrence of cancer (Stanton and Disis, 2015), III) Destroy cancer cells left over from other treatments. The following types of cancer vaccines are being studied: Antigen Vaccines. These vaccines are made from specific proteins or antigens of cancerous cells. Their purpose is to stimulate the immune system to attack cancer cells (Tagliamonte et al., 2014). Whole-Cell Vaccines. A whole-cell vaccine uses the entire cancer cell, not just a specific molecule (antigen), to generate the vaccine. (Keenan and Jaffee, 2012).Dendritic Cell Vaccines. Dendritic cells help the immune system identify abnormal cells, such as cancerous cells. Dendritic cells are grown with cancer cells in the laboratory to produce the vaccine. The vaccine then stimulates the immune system to attack cancer. (Wang et al., 2014; Mastelic-Gavillet et al., 2019). DNA Vaccines. These vaccines are made from DNA fragments of cancer cells. They can be injected into the body to facilitate immune system cells can better respond and kill cancer cells (Gatti-Mays et al., 2017).Other Types of Cancer Vaccines. such as Anti idiotype vaccines. This vaccine stimulates the body to generate antibodies against cancerous cells. An example of an anti-idiotype antibody is Racotumomab or Vaxira (Cancer, 2016). However, conditions and considerations after Corona does not seem to be the same as before. The current pandemic situation has also led to major changes in the pharmaceutical and Vaccine production process and international protocols. Some of the most critical issues that can accelerate the introduction of cancer vaccines are: 1. Typical drug and vaccine development timeline. A typical vaccine needs 5 to 10 years and sometimes longer to design secure funding, and get approval (Figure 1). Less than 10 percent of new drugs, which are entered in the different phases of clinical trials, are advanced to approval by the Food and Drug Administration (FDA)(Cancer, 2020a). However, now the situation is not normal. Dozens of Covid 19 vaccines are starting clinical trials. Some of them use RNA and DNA technology, which delivers the body with missions to produce its antibodies against the virus. There are already at least 254 therapies and 95 vaccines related to Covid-19 being explored. However, it seems that the experiences gained in this pandemic, and advances in technology, may be effective in shortening the production path of other vaccines and drugs and the process of its approval at the national and international levels in the future. In Figure 2, the time course of production of conventional vaccines in comparison with Covid 19 vaccines (Cancer, 2020b) is shown.2. The introduction of messenger RNA (mRNA) technology into the field of prevention and treatment. Over the past decades, this technology has been considered an excellent alternative to conventional vaccination methods. Proper potency and low side effects, the possibility of fast production and relatively low production cost are its advantages. However, until recently, the instability of this molecule has been a major problem in its application. This research was started many years ago by two companies that played a significant role in developing the first Covid vaccines, so BioNTech and Moderna were able to quickly transfer their experience in the field of Covid vaccine development (Pardi et al., 2018; Moderna, 2020). Figure 3 shows how mRNA vaccines work. Bout Pfizer – BioNTech and Moderna mRNA vaccines were more than 90 % effective in preclinical stages. Millions of doses of these two vaccines are currently being injected into eligible individuals worldwide. 3. Considering the use of artificial intelligence in assessing the effectiveness of vaccines. There are always doubts about the effectiveness of the new drug in treating the disease. Once the vaccine is widely available, we will know more about its effectiveness versus it works under carefully controlled scientific testing conditions. Vaccines will continue to be monitored after use. The data collected helps professionals understand how they work in different groups of people (depending on factors such as age, ethnicity, and people with different health conditions) and also the length of protection provided by the vaccine. Artificial intelligence (AI) is an emerging field, which reaches everywhere and not only as a beneficial industrial tool but also as a practical tool in medical science and plays a crucial role in developing the computation vision, risk assessment, diagnostic, prognostic, etc. models in the field of medicine (Amisha et al., 2019). According to the wide range of AI applications in the analysis of different types of data, it can be used in vaccine production, safety assessments, clinical and preclinical studies and Covid 19 vaccines adverse reactions (CDC, 2019). Indeed, most cancer vaccines are therapeutic, rather than prophylactic, and seek to stimulate cell-mediated responses, such as those from CTLs, capable of clearing or reducing tumor burden. There are currently FDA-approved products for helping cancer treatment such as BREYANZI, TECARTUS and YESCARTA for lymphoma, IMLYGIC for melanoma, KYMRIAH for acute lymphoblastic leukemia, and PROVENGE for prostate cancer. Over the past decade, most of BioNTech's activities have been in the field of cancer vaccine design and production for melanoma (two clinical trials), breast cancer (one clinical trial), and the rest concerning viral and veterinary vaccines (two clinical trials). Also Maderno company has been working on Individualized cancer vaccines (one clinical trials), and vaccines for viral infections such as Zika and Influenza and veterinary vaccines (several clinical trials) (Pardi et al., 2018). Therefore, it can be said, mRNA technology that has been the subject of much research into the treatment of cancer has been shifted and rapidly used to produce and use the Covid 19 vaccine. The current pandemic situation has necessitated the acceleration of Covid 19 vaccines and drugs and national and international protocols for their approval. If the currently produced vaccines can continue to be as successful as the preclinical and early phase studies, these changes and evolution have raised hopes for accelerating the use of these technologies and mechanisms in the field of cancer and other diseases vaccines, including HIV and influenza.

Nano magnetism is a subset of physics that examines the magnetic properties of nanometer-sized systems. The magnetic force is caused due to the movement of charged electrical particles. Carriers of magnetic properties in solids are electrons, in which the electrons have a magnetic moment. The physical properties of the magnetic material are dependent on the direction measurement, which is referred to as anisotropy, which is present in many different types. The magnetic nanoparticle production process is carried out chemically in three different phases, after which nanoparticles are synthesized; the surface of these nanoparticles is modified to increase their stability. Magnetic nanoparticles are used because of their unique characteristics in various fields, such as medicine, electronics, food industry, etc. Magnetic nanoparticles have many applications in medicine. These nanoparticles are used in MRI imaging as a contrast agent, which increases the quality of the images. These nanoparticles are widely used in Drug Delivery systems and Hyperthermia for treating cancer and destroying tumors and malignant masses. In the gene delivery system, DNA carrier nanoparticles are transmitted precisely and targeted into the cells of the body. In this method, the healthy gene is placed in an unhealthy gene and the patient’s cells are corrected. Nanoparticles can be used for centrifuges that increase the speed of the field, they can also be used to purge and filter the blood and remove viruses and bacteria from the blood. Nanoparticles are widely used in the manufacture of biosensors. The electrodes in the sensors can be modified using nanoparticles, which reduces the cost of construction. Glucose biosensor is made of SiO2 magnetic particles. Magnetic nanoparticles increase the sensitivity and performance of biosensors. Magnetic nanoparticles in different situations have different magnetic behaviors that are normally in Ferromagnetism or Ferrimagnetism mode.

Spin coated zinc oxide (ZnO)-carbon active porous media on the Fluorine-doped Tin Oxide (FTO) coated glass is presented as an engaged nomination for novel application in enzymatic urea biosensor. Some correlations between the processing parameters spinning speed, spinning duration, volume of solution and ZnO:carbon active ratio and respective thin film characteristics were determined, then uniform ZnO-carbon active the uniform film was achieved at the optimum deposition conditions. FE-SEM was employed to investigate the morphology and hardness of ZnO-carbon active thin film. The FE-SEM image illustrates cavities of a thin film as an efficient transducer area for immobilization of urease enzyme (Urs). Stepwise study of FTO/ZnO-carbon active/Urs biosensor manufacturing was performed by voltammetric and impedimetric techniques. The results revealed a good sensitivity for impedimetric urea retrieval between 8.0-110.0 mg dL-1 with detection limit of 5.4 mg dL-1.

Enzyme immobilization on hydrogels: An overview on methods, interactions, and divers applications.

The JALA Special Issue on Novel Drug Development and Delivery

Chapter 12 - Microbial extraction of micro and nanofibers from plant fibers

Developments in foot-and-mouth disease vaccines

Effect of Surface Curvature and Chemistry on Protein Stability, Adsorption and Aggregation Mithun Radhakrishna Enzyme immobilization has been of great industrial importance because of its use in various applications like bio-fuel cells, bio-sensors, drug delivery and bio-catalytic films. Although research on enzyme immobilization dates back to the 1970’s, it has been only in the past decade that scientists have started to address the problems involved systematically. Most of the previous works on enzyme immobilization have been retrospective in nature i.e enzymes were immobilized on widely used substrates without a compatibility study between the enzyme and the substrate. Consequently, most of the enzymes lost their activity upon immobilization onto these substrates due to many governing factors like protein-surface and inter-protein interactions. These interactions also play a major role biologically in cell signaling, cell adhesion and interprotein interactions specifically is believed to be the major cause for neurodegenerative diseases like Alzheimer’s and Parkinson’s disease. Therefore understanding the role of these forces on proteins is the need of the hour. In my current research, I have mainly focused on two factors a) Surface Curvature b) Surface Chemistry as both of these play a pivotal role in influencing the activity of the enzymes upon immobilization. I study the effect of these factors computationally using a stochastic method known as Monte Carlo simulations. My research work carried out in the frame work of a Hydrophobic-Polar (HP) lattice model for the protein shows that immobilizing enzymes inside moderately hydrophilic or hydrophobic pores results in an enhancement of the enzymatic activity compared to that in the bulk. Our results also indicate that there is an optimal value of surface curvature and hydrophobicity/hydrophilicity where this enhancement of enzymatic activity is highest. Further, our results also show that immobilization of enzymes inside hydrophobic pores of optimal sizes are most effective in mitigating protein-aggregation. These results provide us a rationale to understand the role of chaperonins in protein folding and disaggregation. Our results indicate that strong protein-surface interactions and confinement inducement stability inside pores makes it best suitable for enzyme immobilization.

Characterizing the Janus colloidal particles in AC electric field and a step towards label-free cargo manipulation