Preparation, Characterization, and Applications of Albumin Serum-Based Nanoparticles

Pritimala Sahu^{a, b}, Bhanushree Gupta^b*_,

^a School of Studies in Chemistry, Pt. Ravishankar Shukla University Raipur (C.G.), India 492010

^b Center for Basic Sciences, Pt. Ravishankar Shukla University Raipur (C.G.), India 492010

 

Abstract

Nanoparticles made of albumin have shown great application value in the field of medicine as drug carriers because of their excellent biocompatibility. Albumin-based nanoparticles are generally produced through a desolvation process, which can be strongly affected by pH, temperature, and other desolvating chemicals including ethanol and acetone. In recent years, albumin has been used as a carrier in the diagnosis and treatment of diseases like cancer, HIV, hepatitis, and influenza. Albumin has been shown to be a nontoxic, biocompatible, and biodegradable protein carrier for drug delivery. This review deals with the synthesis of albumin-based nanoparticles and, their characterization through analytical techniques, and their applications. The major characterization techniques include UV-Visible Spectroscopy, Dynamic Light Scattering (DLS), Fourier Transform Infrared Spectroscopy (FT-IR), Field Scanning Electron Microscopy (FESEM), and Transmission Electron Microscopy (TEM). These albumin-based nanoparticles provide an added advantage over other available nanoparticles by being biodegradable and biocompatible. The albumin-based nanoparticles have their applications in medicine and therapeutics, agriculture, cosmetics, and the food industry.

Keywords:  Albumin, Biocompatibility, Nanoparticles, Drug delivery, Antiviral diseases.

1.     Introduction

Proteins are linear polymers of amino acids with a wide range of structural variations and biological activities. They can be classified based on their solubility, chemical structure, shape, and the number of the monomeric unit. Protein solubility is determined by a variety of interactions including protein-protein, protein-water, protein-ions, and ions-water interactions.  Protein-based nanometric structures have been designed for drug delivery systems due to their excellent biocompatibility, biodegradability, and low toxicity (Batchelor et al., 2018). Due to the presence of a number of binding sites in their molecules, protein nanoparticles show a high drug-loading capacity. Covalent bonding, electrostatic attraction, and hydrophobic interaction are involved in drug-loading mechanisms. A variety of proteins have been used in drug delivery systems including albumin, ferritin, transferrin, low-density lipoprotein, and high-density protein (Iqbal et al., 2021). They have also been applied in different targeted therapies like cancer therapy and tumor therapy (Hong et al., 2020).

Albumin proteins are the most common type of proteins used for disease diagnosis and drug delivery/drug carrier systems. Their physical and chemical properties have been highly studied.  Bovine Serum Albumin (BSA) and Human Serum Albumin (HSA) have been used as carrier systems for anticancer drugs (Jimenez-Cruz et al., 2015). There are various types of nanoparticles (NPs) used as drug delivery carriers, including ceramic nanoparticles, magnetic nanoparticles, polymeric nanoparticles, solid-liquid nanoparticles, polymeric micelles, and polymer drugs (Zhou et al., 2013). Many researchers have explored albumin-based nanoparticles, that could accumulate on a cancer cell, to enhance targeting specificity while minimizing side effects to a normal cell (Jahanban-Esfahlan et al., 2016). Nanomedicines can be made multifunctional by adding extra features like the triggered release and targeted distribution (Moghimi et al., 2005). Numerous nanoparticles such as polymer NPs, liposome NPs, inorganic NPs, drug conjugates, hydrogel NPs, viral NPs, etc, have been prepared to be applied in and as various nanoparticle drug delivery systems (Kim et al., 2010, Li et al., 2018 & Liu et al, 2018). Albumins have emerged as excellent carriers and are frequently utilized to encapsulate various therapeutic agents. Albumin-based nanoparticles have shown high binding capacities to many drugs (Wilson et al., 2015).

This study aims to discuss different types of protein and their use in the synthesis of nanoparticles and their application. This review gives an account of a nanoparticle drug delivery system that uses protein as a drug carrier.

 

2.     Albumin Protein

Albumin is a type of protein found in plasma. It plays an important role in our body's physiological processes. It has eight disulfide bonds and a free sulfhydryl group, contributing to its high-water solubility (up to 40% w/v at pH 7.4) (Lei et al., 2021).  Albumin makes up around 50% of all blood proteins and is the main extravascular protein in the blood (35-30 g/L). It is predominantly produced in the liver, at a rate of 0.7 mg/h per gram of liver weight (10–15 g/day) (Iqbal et al., 2021 & Larsen et al., 2016). The highly stable protein albumin, doesn't change structurally after being heated for 10 hours at 60^°C (Jain et al., 2018). In vivo, albumin can be used as a non-specific transport protein to combine with various insoluble organic molecules and inorganic ions to form soluble complexes (Larsen et al., 2016).

 

Types of albumins Protein

a. Ovalbumin

Ovalbumin (OVA) is the most common protein in egg white, and makes up 54-69% of the total protein in egg white (Meng et al., 2022).  It is a highly functional food protein used in the food industry. OVA (Figure 1a) is a typical globulin with a free sulfhydryl group hidden within the hydrophobic core. It is a monomeric phosphoglycoprotein with 47000 Da molecular weight. It consists of 365 amino acids and has an isoelectric point of 4.8. OVA is primarily chosen as a drug carrier owing to its easy availability and low cost (Jimenez-Cruz et al., 2015 & Liu et al., 2021).

 

b. Bovine Serum Albumin

BSA (Figure 1b) is one of the most studied proteins. It has a molecular weight of 69323 Da and an isoelectric point in the water of 4.7 at a temperature of 25^° C (Liu et al., 2021 & Yu et al., 2015).  It is a protein model that is frequently used to explore how proteins accumulate and fibrillate (Karami et al., 2020 & Yu et al., 2015).  BSA has a great potential for the preparation of nano vehicles with possible application in the food industry since it is non-toxic and biodegradable (Jimenez- Cruz et al 2015).  It has three identical homolog domains (I, II, and III) of nine loops. These loops form a triad compound with a large-small-large sequence. Due to the high affinity of BSA it can be used with both hydrophobic and hydrophilic drugs.  Binding interaction of BSA with a drug is shown in Figure 2. BSA is used as a carrier of various drugs like DVH, biotin, etc. because of its high solubility (Karami et al., 2020).

 

 

Figure 1. (a) Ovalbumin (PBD 1OVA)23 (b) Bovine serum albumin (PDB ID 3V03) (Chinnathambi et al., 2015)

 

Figure 2. (a) BSA (PDB ID 3V03), (b) BSA complex with a drug  (Yu et at., 2019)

 

c. Human Serum Albumin

HSA is a globular protein and is mainly found in the human circulatory system. It has a single, typical, non-glycosylated structure (Karami et al., 2020). The human liver produces a majority of this plasma protein, which has 585 amino acids (Yu et al., 2015 & Jimenez-Cruz et al., 2015). It is mostly formed of an alpha-helix structure and lacks a beta-sheet. Lysine amino acid makes up 10% of HSA in total.  HSA (Figure 3) has three identical domains, each consisting of two-sub domains of A and B with the same structural components. The primary protein in plasma, albumin has a concentration of 45 mg/ml and a half-life of 20 days in the blood (Lei et al., 2021). This albumin has no toxicity and is utilized as a drug carrier due to availability and biocompatibility (Batchelor et al., 2018). When HSA is broken down, amino acids provide nutrition to peripheral tissues. The properties of HSA like its preferential uptake in tumor cells, easy availability, biodegradability, and lack of toxicity make it an ideal candidate for drug
delivery (Elzoghby et al., 2012).

 

Figure 3. Human serum Albumin (PDB 1BJ5) (Chinnathambi et al., 2015)

 

3. Protein-Based NPs

 Extensive development and research in the field of nanomedicine gave birth to various design methodologies for protein-based nanoparticles, mainly including desolvation, self-assembly, emulsification, biomineralization and covalent noncovalent interaction. A variety of proteins have been employed to encapsulate or conjugate with chemotherapeutic drugs, imaging agents, radioactive or photoactive agents, and functional ligands for various diagnostic and therapeutic processes. The different methods employed to prepare protein-based NPs generally depend on the properties of drugs and selected proteins, particle size, and the desired application (Tarhini et al 2017). Albumin-based nanoparticles are generally developed through self-assembly, biomineralization, covalent conjugation, or chemical modification methods, depending on specific drug binding sites, cysteine-34, and metal binding site (Jacob et al., 2018). Lipoproteins are naturally represented by lipoprotein nanoparticles (LPN), which are endogenic self-assemblies of protein and lipids. Depending on their hydrodynamic size and surface area, LPNs are divided into four basic groups.  Including high-density LPNs (size 8-12 nm), Low-density LPNs (size 18-25 nm), Very low-density LPNs (size 30-80 nm), and High-density LPNs (size less than 25 nm).

 

4. Significance of Protein-Based nanoparticles

Protein-based nanoparticles are being studied for their applications in pharmaceutical industries and as functional tools due to their biodegradability and low toxicity (Li et al., 2018). Proteins are a preferred alternative to synthesized polymers for applications in the biomedical field because of their safety and the added advantage of biocompatibility. Not only protein nanoparticles can be prepared under mild conditions avoiding the use of toxic chemicals or organic solvents but also their defined primary structure provides multiple possibilities for surface modification (Wang et al., 2014).  In order to be developed as drug delivery systems, protein-based nanoparticles have several advantages like stability, ease of surface modification and particle-size control, low toxicity issues, and biodegradability (Langer et al., 2003). Some advantages of protein-based NPs are presented in Figure 4. Due to their small size, protein-based nanoparticles show high dissolution and can be easily transmitted among cells through endocytosis (Li et al., 2018). Not only these protein nanoparticles are less toxic but they also enhance the release of drugs in biological systems and increase their bioavailability. This factor can be utilized to achieve high efficiency with minimum drug dose and decrease the problem of drug resistance in the human body (Verma et al., 2018).  Since proteins often contain a wide variety of functional groups, they can bind and transport significant amounts of therapeutic compounds by a variety of methods, including electrostatic contacts and hydrophobic interactions, covalent bonds, and interactions (Barchelor et al., 2018). The presence of a number of functional groups in the protein structure allows specific binding of drugs with the nanocarrier and the targeted site in the receptor protein during drug delivery (Ferrara et al., 2021). Protein-based nanoparticles show a non-immunogenic response i.e. they do not stimulate immune response at the receptor site. Also, they show increased cellular uptake as they easily be removed by cells (eg. Vascular epethilialcal) by different mechanisms (Kianfaret al., 2021).

Figure 4. Advantages of protein nanoparticles in drug delivery (Ritz et al., 2015).

5.  Preparation techniques

5.1 Desolvation

This method is used for the preparation of protein-based nanoparticles. In this technique (Figure 5), protein is dissolved in a solution with a pH of 5.5, and ethanol is continuously added dropwise while the solution is stirred until turbid (Jahanban-Esfahlan et al., 2016).  Since the protein component is less soluble in water, its phases separate when ethanol is added to the solution (Lei et al., 2021). The desolvation of protein-based nanoparticles was assessed by Weber and colleagues (Elzoghby et al., 2012).  Weber reported that the minimum glutaraldehyde concentration for producing stable nanoparticles is around 40% with 24 hours of incubation. Particle diameters affected by the pH of the environment (Jahanban-Esfahlan et al., 2016).

 

Figure 5. Preparation of protein-based nanoparticles by desolvation method

5.2 Emulation

The emulation technique is used to create polymeric nanoparticles with protein stabilization using thermal or chemical techniques (Figure 6) (Karami et al., 2020). Using the thermal method, protein nanoparticles are produced by quickly homogenizing the solution containing a protein drop, then heating at 175–180°C for 10 min (Yu et al., 2015). The solution is cooled using ethyl ether to facilitate phase separation during centrifugation. (Lei et al., 2021 & Solanki et al., 2021).  In the chemical method, the aqueous protein solution is stabilized by emulsification at 25°C into the oil. The aqueous protein solution is then denatured using formaldehyde suspension (kim et al., 2010 & Liu et al., 2018).

Figure 6. Preparation of protein-based nanoparticles by emulation method

 

5.3 Thermal gelation

The thermal gelation technique is used to synthesize gel from a solution by heating (Figure 7). This process involves the heat-induced unfolding of native protein structure followed by protein-protein interactions involving hydrogen ions, electrostatic forces, and hydrophobic bonds (Yedomon et al 2013). Thermal gelation often involves mixing lysozyme and albumin solution at pH 5.3 to create nanoparticles. The pH is then set to 10.3 and the produced solution is heated again (Lei et al., 2021 & Yu et al., 2015).

Figure 7. Preparation of protein-based nanoparticles by thermal gelation.

 

5.4 Nanoparticle albumin-bound (NAB) technology

NAB technology was developed by an American bioscience company. In this method (Figure 8), a drug is combined with a selected albumin protein in an aqueous solution to create protein nanoparticles that range in size from 100 to 200 nm (Moghimi et al., 2005 & Jimenez_Cruz et al., 2015). The NP NAB technology was developed to make NPs that are safe and appropriate for intravenous administration of anticancer drugs with limited solubility.

Figure 8. Preparation of albumin-Based nanoparticles by NAB technique method.

 

5.5 Self-Assembly

In this technique, protein nanoparticles involve an arrangement of protein units into ordered structures by linking the monomers. This method (Figure 9) of producing self-assembly protein nanoparticles (SAPNs) involves increasing the hydrophobic power of protein by adding lipophilic drugs and decreasing amino groups from the surface of the proteins (Tarhini et al., 2017).

Figure 9. Preparation of albumin-Based nanoparticles by Self-assembly method

 

5.6 Nano spray drying

Spray drying is a well-established method commonly used in the pharmaceutical industry to produce dry powder from a liquid phase. Nanoparticles in liquid samples can be processed using nano spray drying (Figure 10). The nozzle sprays liquid samples into chambers that contain hot nitrogen and carbon dioxide gas flowing in the same direction. (Vandervoort, Ludwig 2002, & Ahmed et al., 2012). Nanoparticles are collected using an electrode located at the bottom of the chamber. These electrodes provide an electrostatic charge to the sprayed droplets as they go toward the chamber's bottom. This is a step-by-step process as well as a fast and cost-effective way of producing small-scale protein particles. Lee et al. employed the innovative Nano Spray Dryer B-90 and Tween 80 as a surfactant to create BSA nanoparticles in a single step in a practical manner. (equipped with a vibrating mesh spray technology and an electrostatic particle collector) (Lee et al., 2011). When the surfactant was added, the particles' form shifted to a spherical one, stabilizing the nanoparticles. Optimized production of smooth spherical nanoparticles (median size:460±10nm and yield:72±4%) was achieved using the 4μm spray mesh at 0.1%w/v BSA concentration, 0.05%w/v surfactant concentration, drying flow rate of 150L/min and inlet temperature of 120°C (Lee et al., 2011).

Figure 10. Preparation of albumin-Based nanoparticles by Nano spray drying method.

 

5.7 Complex coacervation method

In the coacervation process (Figure 11), when a salt is added to a protein solution, liquid-liquid phase separation occurs, giving rise to a polymer-rich dense phase at the bottom and a transparent solution above resulting in the desired NPs (Lu et al., 2004 & Elzoghby et al., 2015).  Proteins include several charged functional groups and are amphoteric; they can be made cationic or anionic by varying several variables, including the protein's pH. Other polymeric electrolytes can engage in electrostatic interactions with the charged protein. Proteins and other polymers can interact electrostatically at pH-dependent levels to create stable, biocompatible nanoparticles and coacervates that enable the regulated transfer of DNA and bioactive therapies. (Ahmed et al., 2012).

Figure 11. Preparation of albumin-Based nanoparticles by Complex coacervation method method.

5.8 Ionotropic gelation

Calvo et al. (1997) first reported this technique and has been widely examined and developed. The method utilizes the electrostatic interaction between the amine group of protein/ biopolymer and a negatively charged group of polyanion such as tripolyphosphate (Figure 12). Protein can be dissolved in an organic solvent of stabilizing agents, such as poloxamer. Polyanion was then added, and nanoparticles were formed spontaneously under mechanical stirring at room temperature. The size and surface charge of particles can be modified by changing the ratio of proteins/biopolymers to the stabilizer. A general increase in particle compactness and size was observed by increasing the proteins/biopolymer concentration and increasing the polymer to polyanion ratio (Diyva et al., 2018 & Jonassen et al., 2012). They also reported that the smaller particle size found in the presence of sodium chloride made nanoparticles dispersed in saline solution more stable. This is because a univalent salt like sodium chloride when added to the solvent screens out to the electrostatic repulsion between the positively charged amine groups on the proteins/biopolymer backbone. This will increase the flexibility of the polymer chains in solution and thus increase its stability (Ilium 1998). The advantages and disadvantages of different preparation techniques used in the synthesis of protein-based nanoparticles are discussed in Table 1.

 Figure 12.  Preparation of protein/biopolymer-Based nanoparticles by Ionotropic gelation method.

Table 1.  Advantages and disadvantages of preparation techniques of protein-based nanoparticles (Hong et al., 2020).

 

6. Characterization techniques

 6.1 Dynamic Light Scattering (DLS) system

Dynamic light scattering (DLS) determines the particle diameter with the aid of Brownian motion and light scattering properties. He-Ne laser operation at 90° angle are used in dynamic light scattering (DLS) equipment with a 4 mW to detect the average size and PDI (Yedomon et al., 2013 & Elzoghby et al., 2015). DLS is based on scattered light interacting with a sample which provides information about the size of nanoparticles, PDI, and nanoparticle’s stability (Verma et al., 2015). The PDI is a measure of a sample’s particle size distribution. It ranges from 0 (monodispersed) to 1 (broad distribution) (Al-jawad et al., 2018).

 

6.2 UV-Visible Spectrophotometer

UV-Vis Spectroscopy is a type of electronic microscopy. It is based on the principle of Beer- Lambert law and is used for quantitative analysis of the sample. According to law, when the monochromatic beam of light passes through the homogenous absorbing medium, then the absorbance of the sample is in direct proportion to the path length (l) and concentration (c) of absorbing moieties in the sample at a particular wavelength. Deuterium tungsten lamps can be used as a light source for UV-Vis Spectroscopy (Jimenez-Cruz et al., 2015). In the range of 200-600nm, UV-Visible spectrophotometer measurements of albumin-based nanoparticles are made at 25⁰C in the range of 200-600 nm (Jahanban- Esfahlan et al., 2016).

 

6.3 Field Scanning Electron Microscopy (FESEM) study-

It is a high-resolution image technique focused on the high energy electron beam (Verma et al., 2015). The secondary electron emitted during this interaction provides the insight about the geometry, topography and composition of the sample under investigation The sample must be electronically grounded and conducting in nature for the imaging. the non conductive specimens are often coated with the thin layer of metals, mainly platinum, tungsten, gold, chromium and graphite, that prevents the accumulation of the negative charge on the surface. At room temperature, the protein nanoparticles can be dispersed using sonication and then subjected to analysis at a voltage of 5–20 kV (Jahanban- Esfahlan et al., 2016).

 

6.4 Transmission Electron Microscopy (TEM)

It is a high-resolution imaging technique that involves electrons with very short wavelengths for imaging. TEM images for protein nanoparticles can be captured at an accelerating voltage of 100 kV (Karami et al., 2020). Initially the prepared samples need to be stained with a 2% solution of ammonium molybdate that had been filtered through a 0.45µm millipore filter (Yedomon et al., 2013). The nanoparticle suspension can then be applied to the carbon-coated copper grid, and any excess solution should be removed with filter paper.

 

6.5 Fourier Transform Infrared (FTIR) Spectroscopy

It is based on the vibrational transition that can be the determination of the functional group associated with the nanoparticles (Elzoghby et al., 2015 & Jahanban-Esfahlan et al., 2016).  FT-IR spectroscopy is used to detect and record the spectra throughout a 400–4000 cm^-1 wavelength range.  The region between 1500-400 cm^-1 is referred to as a fingerprint region and 4000-1600 cm^-1 is regarded as a functional group region. Only the molecules with net dipole considered are IR active. The functionalization of nanoparticles can easily determine the bond strength, types of bonds, and structure of molecules (Verma et al., 2020).

 

6.6 X-Ray diffraction (XRD)

X-Ray Diffraction technique is a significant tool used to identify the nanoparticles' structure. It is a non-destructive, non-contact technique that can provide extensive information on the chemical and physical characteristics of the sample, but most crucially about the crystal structure, including crystallite size, crystallite strain, orientation, and defect. It has a quantitative approach (Jahanban-Esfahlan et al., 2016 & Karami et al., 2020). It can be determined using Bragg’s law [Eq. (1)]

nλ = 2dsinθ                (1)

where, n= order of reflection, λ = x-ray wavelength, d= interplanar distance, θ= angle between incident beam and lattice plane.

 

6.7 Atomic Force Microscopy (AFM)

 Atomic Force Microscopy is a type of scanning probe microscopy, it is a non-destructive technique, that can be used in the characterization of various surface measurements such as texture and roughness by provideding 3D topographic images of nanoparticles (Verma et al., 2015). AFM can be operated in majorly three modes such as contact mode, non-contact mode, and tapping mode (Elzoghby et al., 2012).

 

7.  Encapsulation and drug loading Efficiency of Albumin-based nanoparticles

The effectiveness of drug encapsulation plays an important role in the selection of a drug-delivery vehicle. This efficiency can be calculated in percentage using Equations [Eq. (2) & (3)] (Subia, Kundu 2012).

 

Encapsulation efficiency =  × 100 %     (2)

 

Drug Loading efficiency = × 100 %              (3)

 

The concentration of the free, nonentrapped medication is ascertained by producing a calibration curve of varying amounts of the entrapped drug using UV-visible spectroscopy (Figure 13). K.P.Singh and group evaluated the encapsulation efficiency of albendazole-encapsulated Liposome and PEG-coated Liposome and found the values to be 72% and 81% respectively (Panwar et al., 2010).

 

Figure 13  Method of drug loading

 

8. In Vitro release studies

The drug release rate is an important factor in selecting a nanocarrier for drug delivery. The in vitro studies for drug release from encapsulated protein nanoparticles can be performed by dialysis method. K.P. Singh and group have studied and compared the in vitro release behavior of free and encapsulated albendazole. Encapsulation was done by both conventional and PEG-coated Liposome systems (Panwar et al., 2010).  S.C. Kundu and group studied the in vitro release behavior of methotrexate from silk fibroin- Albumin nanocarriers and found that 85% drug was released after 12 days (Subia, Kundu 2012). The encapsulation, loading, and release of drugs are highly effected by size, change, and hydrophobicity of the carrier system.

 

9. Applications of protein-based nanoparticles in drug delivery

Protein-based nanoparticles are being used as drug-delivery systems in the field of medicine. The surface of protein nanoparticles can be easily functionalized in addition to being biocompatible and biodegradable due to their specified basic structure. Also, charged proteins can assist drug loading through electrostatic interactions. The scopes of the protein nanocarrier systems have been investigated both in vitro and in vivo to encapsulate several medicinal compounds. Albumin proteins are used as a carrier in the treatment of cancer, diabetes, etc. Several albumin-based drug delivery systems are commercially available like  abraxane, levmir and victoza in the treatment of diabetes, (Frates et al., 2018) ozoralizumab in the treatment of rheumatoid arthritis and albuferon in the treatment of hepatitis C (Elsadek et al., 2012). Satya Prakash and group have developed and assessed HSA  nanoparticle for the targeted delivery of milrinone for  the treatment of congestive Heart failure (Sarfaraz et al., 2021).  Liposomes have also been used for a number of clinical applications.

 

9.1 Role of protein-based nanoparticles in cancer treatment

Cancer is one of the terrible diseases that the world has been dealing with for years. Albumin nanospheres have been used to entrap chemotherapy drugs such 5-fluorouracil, doxorubicin, and methotrexate, and their properties have been thoroughly investigated (Lomis et al., 2021). Protein nanoparticles have been employed in different types of cancer therapies.

 

Chemotherapy 

One of the most popular forms of cancer treatment, chemotherapy uses chemotherapeutic medications to destroy cancer and achieve the desired therapeutic outcome. Chemotherapy frequently has a long list of side effects because of the non-specific toxicity of the drugs (Jose et al., 2014 & Lei et al., 2021) The albumin-based nanoparticles usually have the following advantages over conventional drug delivery techniques for the delivery of anticancer medicines (Zhou et al., 2013 & Yu et al., 2015).

 

Gene therapy

Exogenetic genes are inserted into target cells during gene therapy to correct defective genes and express desirable proteins in order to achieve therapeutic goals. Gene therapy has significantly advanced cancer treatment over the last few decades. However, exogenous nucleic acids can be quickly eliminated when supplied through in vivo delivery (Joye et al., 2014). Due to their versatility, the use of biopolymeric nanocarriers like protein nanoparticles in drug or gene delivery systems made it simple to reach specific target areas (Chen et al., 2018, & Jeslin et al., 2018). Before the drug or gene can be delivered, the nanocarriers carrying the drug or gene must get past a variety of biological obstacles and enter the bloodstream. The biocompatibility of protein nanoparticles makes them advantageous over other synthetic polymeric nanoparticles. Different tactics, including active targeting, passive targeting, and stimuli-modulated drug release can be used to accomplish gene therapy through protein nanoparticles (Jeslin et al., 2018 & Nitta et al., 2013).

 

9.2 Role of Protein-Based nanoparticles in delivery of antiviral drugs

Protein-based nanoparticles can be applied to encapsulate and deliver anti-viral drugs. J. M. Irache and group have studied intravitreal delivery of two different anti-cytomegaloviral drugs ganciclovir and formivisen (Wu et al., 2019).  Ocular drug delivery systems have also been developed with BSA nanoparticles. These systems have been studied for ocular delivery of Acyclovir, an antiviral compound (Irache et al., 2005 & Suwannoi et al., 2017).

 

10. Conclusion

Protein-based nanoparticles are a better alternative to metallic nanoparticles and other synthetic polymeric nanoparticles due to their properties like biocompatibility, biodegradability, ease of surface modification, ease of particle size control, non-immunogenic response, increased cellular uptake and targeted binding to the receptor site. Using biopolymeric nanoparticles is a great and economically viable approach for the targeted delivery of drugs for treatment of various diseases. Studies have shown that using protein nanoparticles as nanocarriers increases the stability and effectiveness of drugs while reducing their toxicity. Such protein-based nanocarriers have found various applications in the field of medicine as drug delivery systems, especially in the case of cancer treatment. However, their applications in the treatment of other life-threatening diseases need to be explored further.

 

11. Future perspective 

Albumin-based nanocarriers are predicted to play an important role in combined therapy and theranostic in the future. Considering the harmful side-effects of commercially available cosmetics and skin-care products, the potential of protein nanoparticles must also be explored in this category. The potential of protein-based nanoparticles can also be explored to enhance the quality and quantity of products in the field of agriculture, food, cosmetics, etc. The application of structurally designed biopolymer particles in the food industry is still in the initial stages of research. Food companies are looking for ways to make the healthiness of their products without compromising their quality or the way consumers perceive them. Toxicological studies should also be done to make sure that the new technologies are safe for wide use in the food industry.

 

12. Acknowledgment

We are thankful to the Head of Department, School of Studies in Chemistry and Director, Center for Basic Sciences, Pt. Ravishankar Shukla University, Raipur for providing the research facilities. The authors are grateful to the Department of Science & Technology, Government of India for providing financial assistance (DST/INSPIRE Fellowship/2022/IF220056).

 

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