Stabilization challenges and aggregation in protein-based therapeutics in the pharmaceutical industry.

Protein-based therapeutics have revolutionized the pharmaceutical industry and become vital components in the development of future therapeutics. They offer several advantages over traditional small molecule drugs, including high affinity, potency and specificity, while demonstrating low toxicity and minimal adverse effects. However, the development and manufacturing processes of protein-based therapeutics presents challenges related to protein folding, purification, stability and immunogenicity that should be addressed. These proteins, like other biological molecules, are prone to chemical and physical instabilities. The stability of protein-based drugs throughout the entire manufacturing, storage and delivery process is essential. The occurrence of structural instability resulting from misfolding, unfolding, and modifications, as well as aggregation, poses a significant risk to the efficacy of these drugs, overshadowing their promising attributes. Gaining insight into structural alterations caused by aggregation and their impact on immunogenicity is vital for the advancement and refinement of protein therapeutics. Hence, in this review, we have discussed some features of protein aggregation during production, formulation and storage as well as stabilization strategies in protein engineering and computational methods to prevent aggregation.

Impact of Enterococcus italicus ONU547 on the growth and acclimatization of micropropagated Rubus fruticosus L. and Paulownia tomentosa Steud. plants to ex vitro conditions.

Clonal micropropagation is an effective method for plant reproduction, applicable in both scientific and industrial domains. However, a significant number of microclones are lost during the ex vitro acclimatization process. To address this, the introduction of beneficial microorganisms into the rhizosphere of micropropagated plants could have a positive effect on the survival rates and external characteristics of acclimatized plantlets. The aim of this study was to determine the protective and growth-promoting potential of Enterococcus italicus ONU547 and its effect on micropropagated plants during acclimatization. The antagonistic activity of the bacteria was determined using the agar block method. Lepidium sativum L. seeds were inoculated with bacterial suspensions at concentrations of 106, 107, and 108 CFU/ml. Subsequently, the roots of the microclones were treated with suspensions of 106 and 107 CFU/ml, and biometric characteristics were measured. The results demonstrated antagonistic properties against various phytopathogenic fungi, including Aspergillus niger, Cladosporium cladosporioides, Alternaria alternata, Alternaria tenuissima, Rhizoctonia cerealis, Penicillium expansum, and Paecilomyces variotii. Inoculation of L. sativum L. seeds resulted in improved germination rates, increased root numbers, and enhanced root and shoot lengths. Similarly, the effects of the studied bacteria on Rubus fruticosus L. and Paulownia tomentosa Steud. during the acclimatization stage led to higher survival rates, increased shoot lengths, greater node numbers, and larger leaf areas. A concentration of 107 CFU/ml was identified as optimal for inoculating the microclones. The findings indicate that E. italicus ONU547 holds promise for the inoculation of micropropagated plants during the acclimatization process. Further research is recommended to establish the specific interaction mechanisms between these bacteria and plants.

Probiotic and technological characterization of selected Lactobacillus strains isolated from different egyptian cheeses.

BACKGROUND: Fresh milk and natural environmental conditions are used to produce traditional cheeses. Such cheeses are produced by dozens of different types of microbes. Non-starter lactobacilli are the most responsible genus of lactic acid bacteria exhibiting key technological and health promoting traits. The purpose of this study is to isolate Lactobacillus bacteria from conventional Egyptian cheeses and analyse their probiotic potential and technological properties. RESULTS: Lactobacillus isolates (33 isolates) were isolated from different Egyptian cheeses. Our results revealed that 18.18% of the isolates were fast-acidifying, 30.3% were medium-acidifying and 51.5% were slow-acidifying isolates. The results of autolytic activity showed that 24.3% of the isolates were good autolysis, 33.3% were fair autolysis, while 42.4% were poor autolysis. Fifteen isolates produced exopolysaccharides, while 9 isolates exhibited antimicrobial activities against Lactobacillus bulgaricus 340. All the isolates were resistant to pH 3 for 3 h except isolate No. 15 (MR4). The growth rate of the isolates ranged from 42.25 to 85.25% at 0.3% bile salts after 3 h of incubation. The surviving percentage of the Lactobacillus isolates decreased with increasing incubation time or the percentage of bile salts greater than 0.3%. All the isolates grew after incubation in artificial gastric and intestinal fluids. The auto-aggregation of 15 isolates ranged from 43.13 to 72.77%. Lacticaseibacillus paracasei BD3, Lactiplantibacillus plantarum BR4 and Limosilactobacillus fermentum MR2 were sensitive to the majority of the tested antibiotics and showed good bile salt hydrolase activity. CONCLUSION: L. paracasei BD3, L. plantarum BR4 and L. fermentum MR2 were isolated from Egyptian cheeses and showed probiotic and technological characterization, which are valuable for their practical application as starters, adjunct and protective cultures in cheese making.

Modulation of cytotoxic amyloid fibrillation and mitochondrial damage of α-synuclein by catechols mediated conformational changes.

The interplay between α-synuclein (α-syn) and catechols plays a central role in Parkinson's disease. This may be related to the modulating effects of catechols on the various aspects of α-syn fibrillization. Some of these effects may be attributed to the membrane-binding properties of the protein. In this work, we compare the effect of some catechols, including dopamine, epinephrine, DOPAL, and levodopa in micromolar concentrations, on the in vitro cytotoxicity of α-syn fibrils on human neuroblastoma SH-SY5Y cells. The study was followed by comparing the interactions of resulting structures with rat brain mitochondria used as an in vitro biological model. The obtained results demonstrate that catechols-induced structures have lost their cytotoxicity mimicking apoptotic cell death mediated by α-syn aggregates in different proportions. Moreover, α-syn fibrils-induced mitochondrial dysfunction, evaluated by a range of biochemical assays, was modulated by catechols-modified α-syn oligomers in different manners, as levodopa and DOPAL demonstrated the maximal and minimal effects, respectively. The plausible mechanism causing the inhibition of α-syn cytotoxic fibrillization and mitochondrial dysfunction by catechols is discussed. Taken together, we propose that catechols can prevent the cytotoxic assembly of α-syn and its destructive effects on mitochondria at various stages, suggesting that decreased levels of catechols in dopaminergic neurons might accelerate the α-syn cytotoxicity and mitochondrial dysfunction implicating Parkinson's disease.

Bivalent metal ions induce formation of α-synuclein fibril polymorphs with different cytotoxicities.

α-Synuclein (α-Syn) aggregates are key components of intracellular inclusion bodies characteristic of Parkinson's disease (PD) and other synucleinopathies. Metal ions have been considered as the important etiological factors in PD since their interactions with α-Syn alter the kinetics of fibrillation. In the present study, we have systematically explored the effects of Zn2+, Cu2+, Ca2+, and Mg2+ cations on α-Syn fibril formation. Specifically, we determined fibrillation kinetics, size, morphology, and secondary structure of the fibrils and their cytotoxic activity. While all cations accelerate fibrillation, we observed distinct effects of the different ions. For example, Zn2+ induced fibrillation by lower tlag and higher kapp and formation of shorter fibrils, while Ca2+ ions lead to formation of longer fibrils, as evidenced by dynamic light scattering and atomic force microscopy studies. Additionally, the morphology of formed fibrils was different. Circular dichroism and attenuated total reflection-Fourier transform infrared spectroscopies revealed higher contents of ß-sheets in fibrils. Interestingly, cell viability studies indicated nontoxicity of α-Syn fibrils formed in the presence of Zn2+ ions, while the fibrils formed in the presence of Cu2+, Ca2+, and Mg2+ were cytotoxic. Our results revealed that α-Syn fibrils formed in the presence of different divalent cations have distinct structural and cytotoxic features.

Thermal stability enhancement: Fundamental concepts of protein engineering strategies to manipulate the flexible structure.

Increasing the temperature by just a few degrees may lead to structural perturbation or unfolding of the protein and consequent loss of function. The concepts of flexibility and rigidity are fundamental for understanding the relationships between function, structure and stability. Protein unfolding can often be triggered by thermal fluctuations with flexible residues usually on the protein surface. Therefore, identification and knowledge of the effect of modification to flexible regions in protein structures are required for efficient protein engineering and the rational design of thermally stable proteins. The most flexible regions in protein are loops, hence their rigidification is one of the effective strategies for increasing thermal stability. Directed evolution or rational design by computational prediction can also lead to the generation of thermally stable proteins. Computational protein design has been improved significantly in recent years and has successfully produced de novo stable backbone structures with optimized sequences and functions. This review discusses intramolecular and intermolecular interactions that determine the protein structure, and the strategies utilized in the mutagenesis of mesophilic proteins to stabilize and improve the functional characteristics of biocatalysts by describing efficient techniques and strategies to rigidify flexible loops at appropriate positions in the structure of the protein.

Allergenicity of Fermented Foods: Emphasis on Seeds Protein-Based Products.

Food allergy is an IgE-mediated abnormal response to otherwise harmless food proteins, affecting between 5% and 10% of the world preschool children population and 1% to 5% adults. Several physical, chemical, and biotechnological approaches have been used to reduce the allergenicity of food allergens. Fermentation processes that contribute to technological and desirable changes in taste, flavor, digestibility, and texture of food products constitute one of these approaches. Lactic acid bacteria (LAB), used as starter cultures in dairy products, are a subject of increasing interest in fermentation of plant proteins. However, the studies designed to assess the impact of LAB on reduction of allergenicity of seed proteins are at an early stage. This review presents the current knowledge on food fermentation, with a focus on seed proteins that are increasingly used as ingredients, and its impacts on food potential allergenicity.

Efficiency of milk proteins in eliminating practical limitations of ß-carotene in hydrated polar solution.

The objective of this work was to study ß-carotene functionalities (color and antioxidant activity) and practical limitations (aggregate formation, poor solubility and low stability) when included in the aqueous systems containing milk proteins. According to the results, self-association constant of ß-carotene in the presence of casein is 1.7-fold of that calculated for WPI. Casein and WPI were capable of conserving ß-carotene against chemical oxidation up to 15 and 12%, respectively, at 1:5 M ratio of ß-carotene to protein. While, WPI reduced its photodegradation quantum yield from 0.03 to 0.012 compared to 0.017 obtained for casein. A 2.7- and 3.6-fold enhancement in ß-carotene solubility was observed in the presence of 1.5 mg/mL of casein and WPI, respectively. The study of ß-carotene interaction with proteins showed, on the one hand, a negative effect on electron transfer and, on the other hand, improved hydrogen transfer to the radical species in the solution.

Helicobacter pylori and Its Antibiotic Heteroresistance: A Neglected Issue in Published Guidelines.

"Heteroresistance" is a widely applied term that characterizes most of the multidrug-resistant microorganisms. In microbiological practice, the word "heteroresistance" indicates diverse responses to specific antibiotics by bacterial subpopulations in the same patient. These resistant subpopulations of heteroresistant strains do not respond to antibiotic therapy in vitro or in vivo. Presently, there is no standard protocol available for the treatment of infections caused by heteroresistant Helicobacter pylori in clinical settings, at least according to recent guidelines. Thus, there is a definite need to open a new discussion on how to recognize, how to screen, and how to eliminate those problematic strains in clinical and environmental samples. Since there is great interest in developing new strategies to improve the eradication rate of anti-H. pylori treatments, the presence of heteroresistant strains/clones among clinical isolates of the bacteria should be taken into account. Indeed, increased knowledge of gastroenterologists about the existence of heteroresistance phenomena is highly required. Moreover, the accurate breakpoints should be examined/determined in order to have a solid statement of heteroresistance among the H. pylori isolates. The primary definition of heteroresistance was about coexistence of both resistant and susceptible isolates at the similar gastric microniche at once, while we think that it can be happened subsequently as well. The new guidelines should include a personalized aspect in the standard protocol to select a precise, effective antibiotic therapy for infected patients and also address the problems of regional antibiotic susceptibility profiles.

Combined microwave processing and enzymatic proteolysis of bovine whey proteins: the impact on bovine ß-lactoglobulin allergenicity.

The main aim of this study was to develop a continuous microwave treatment system of whey proteins and then apply this process at 37 °C, 50 °C, 65 °C and 70 °C to achieve pepsinolysis and produce extensively hydrolysed bovine whey protein hydrolysates with low allergenic properties. The microwave process was compared to a conventional thermal treatment with similar temperature set points. Both processes were deeply analysed in terms of the thermal kinetics and operating conditions. The pepsin hydrolysates obtained by the continuous microwave treatment and conventional heating were characterized by SDS-PAGE and RP-HPLC. The allergenicity of the whey protein hydrolysates was explored using a human IgE sensitized rat basophil leukaemia cell assay. The results indicate that extensively hydrolysed whey protein hydrolysates were obtained by microwave only at 65 °C and in a shorter time compared with the conventional thermal treatment. In the same temperature conditions under conventional heating, ß-lactoglobulin was resistant to pepsinolysis, and 37% of it remained intact. As demonstrated by an in vitro degranulation assay using specific human IgE-sensitized rat basophils, the extensively hydrolysed whey protein obtained by microwave showed maximum degranulation values of 6.53% compared to those of the native whey protein isolate (45.97%) and hence elicited no more allergenic reactions in basophils. This work emphasizes the potential industrial use of microwave heating specific to milk protein processing to reduce their allergenicity and improve their end-use properties.

A health concern regarding the protein corona, aggregation and disaggregation.

Nanoparticle (NP)-protein complexes exhibit the "correct identity" of NP in biological media. Therefore, protein-NP interactions should be closely explored to understand and modulate the nature of NPs in medical implementations. This review focuses mainly on the physicochemical parameters such as dimension, surface chemistry, morphology of NPs, and influence of pH on the formation of protein corona and conformational changes of adsorbed proteins by different kinds of techniques. Also, the impact of protein corona on the colloidal stability of NPs is discussed. Uncontrolled protein attachment on NPs may bring unwanted impacts such as protein denaturation and aggregation. In contrast, controlled protein adsorption by optimal concentration, size, pH, and surface modification of NPs may result in potential implementation of NPs as therapeutic agents especially for disaggregation of amyloid fibrils. Also, the effect of NPs-protein corona on reducing the cytotoxicity and clinical implications such as drug delivery, cancer therapy, imaging and diagnosis will be discussed. Validated correlative physicochemical parameters for NP-protein corona formation frequently derived from protein corona fingerprints of NPs which are more valid than the parameters obtained only on the base of NP features. This review may provide useful information regarding the potency as well as the adverse effects of NPs to predict their behavior in vivo.

Characterization of the bacteriocin produced by Enterococcus italicus ONU547 isolated from Thai fermented cabbage.

Seventy-eight isolates of lactic acid bacteria from Ukraine and Thailand were screened for bacteriocinogenic activity against indicator strain Lactobacillus sakei subsp. sakei JCM 1157. One isolate showed an antagonistic activity of cell-free supernatant eliminated after the treatment with Proteinase K. Based on 16S rRNA gene sequence, this isolate was identified as Enterococcus italicus. Bacteriocin produced by this strain showed antimicrobial activity against L. sakei subsp. sakei JCM 1157, Brochothrix thermosphacta DSMZ 20171, and Listeria ivanovii subsp. ivanovii DSMZ 20750 in agar well diffusion assay. This bacteriocin was cationic and hydrophobic. The partially purified bacteriocin was thermostable, while heating of cell-free supernatant increased its activity more than twofold. Molecular mass of the partially purified bacteriocin as determined by SDS-PAGE differed from enterocin A and B previously known for E. italicus. Concentrated bacteriocin decreased the level of biofilm formation in L. sakei subsp. sakei JCM 1157 and Pseudomonas aeruginosa PAO1 in 52.5 and 48.0%, respectively (p < 0.05). We suggest that the studied bacteriocin could be a perspective antibiofilm agent in food conservation and medicine.

Binding of ß-carotene to whey proteins: Multi-spectroscopic techniques and docking studies.

The objective of this work was to study molecular binding between ß-carotene (ß-C) and whey protein isolate (WPI) as a function of pH (4-9), temperature (15, 25, and 35 °C), and NaCl concentration (0-0.25 M) using spectroscopic techniques and docking studies. The fluorescence quenching data showed that binding affinity increased with pH, temperature and ionic strength. The binding was entropy driven and involved mostly hydrophobic interactions. Three major whey proteins including ß-lactoglobulin (ß-Lg), α-lactalbumin (α-Lac), and bovine serum albumin (BSA) were bound to ß-C with overall binding constant values of 1.31 × 107, 1.80 × 104, and 4.51 × 104 M-1, respectively. A single class of binding sites for ß-C on whey fractions was recognized using Job's method. Docking results revealed ß-C was bound to the subdomain IIA of BSA, the residues of aromatic cluster II in α-lactalbumin and into the calyx of ß-lactoglobulin resulting in conformational changes in the secondary and tertiary structures of proteins.

A biophysical study on the mechanism of interactions of DOX or PTX with α-lactalbumin as a delivery carrier.

Doxorubicin and paclitaxel, two hydrophobic chemotherapeutic agents, are used in cancer therapies. Presence of hydrophobic patches and a flexible fold could probably make α-Lactalbumin a suitable carrier for hydrophobic drugs. In the present study, a variety of thermodynamic, spectroscopic, computational, and cellular techniques were applied to assess α-lactalbumin potential as a carrier for doxorubicin and paclitaxel. According to isothermal titration calorimetry data, the interaction between α-lactalbumin and doxorubicin or paclitaxel is spontaneous and the K (M-1) value for the interaction of α-lactalbumin and paclitaxel is higher than that for doxorubicin. Differential scanning calorimetry and anisotropy results indicated formation of α-lactalbumin complexes with doxorubicin or paclitaxel. Furthermore, molecular docking and dynamic studies revealed that TRPs are not involved in α-Lac's interaction with Doxorubicin while TRP 60 interacts with paclitaxel. Based on Pace analysis to determine protein thermal stability, doxorubicin and paclitaxel induced higher and lower thermal stability in α-lactalbumin, respectively. Besides, fluorescence lifetime measurements reflected that the interaction between α-lactalbumin with doxorubicin or paclitaxel was of static nature. Therefore, the authors hypothesized that α-lactalbumin could serve as a carrier for doxorubicin and paclitaxel by reducing cytotoxicity and apoptosis which was demonstrated during our in vitro cell studies.

ß-Cyclodextrin-Modified Magnetic Nanoparticles Immobilized on Sepharose Surface Provide an Effective Matrix for Protein Refolding.

In this article, we propose an impressive and facile strategy to improve protein refolding using solid phase artificial molecular chaperones consisting of the surface-functionalized magnetic nanoparticles. Specifically, monotosyl-ß-cyclodextrin connected to the surface of 3-aminopropyltriethoxysilane (APES)-modified magnetic nanoparticles is immobilized on the sepharose surface to promote interaction with exposed hydrophobic surfaces of partially folded (intermediates) and unfolded states of proteins. Their efficiencies were investigated by circular dichroism spectroscopy and photoluminescence spectroscopy of the protein. Although the mechanism of this method is based on principles of hydrophobic chromatography, this system is not only purging the native protein from inactive inclusion bodies but also improving the protein refolding process. We chose ß-cyclodextrin (ß-CD) considering multiple reports in the literature about its efficiency in protein refolding and its biocompatibility. To increase the surface area/volume ratio of the sepharose surface by nanoparticles, more ß-CD molecules are connected to the sepharose surface to make a better interaction with proteins. We suppose that proteins are isolated in the nanospace created by bound cyclodextrins on the resin surface so intermolecular interactions are reduced. The architecture of nanoparticles was characterized by Fourier transform infrared spectra, X-ray diffraction, scanning electron microscopy images, energy dispersive X-ray spectroscopy, nuclear magnetic resonance (1H NMR and 13C NMR), and dynamic light scattering.

Thermodynamic, crystallographic and computational studies of non-mammalian fatty acid binding to bovine ß-Lactoglobulin.

The milk protein ß-lactoglobulin has been widely studied since its discovery, both as a purified protein and in mixtures with other milk proteins, where its effect on the processing properties is of importance to the dairy industry. The protein can bind a variety of small hydrophobic molecules, which may allow its use as an oral delivery vehicle. In the present study we have examined the binding of odd-numbered fatty acids by isothermal calorimetry (ITC), X-ray crystallography and computer modelling to provide a clearer picture of the extent and variability of the central binding pocket. The Kd values for the fatty acids C13, C15, C16, C17 and C19 as determined by ITC are 1.93, 2.91, 3.05, 4.11 and 8.67 × 10-7 M, respectively. The molecular structures revealed the ligands bound in the central cavity with generally well ordered lipophilic tails but significant positional variation at the carboxyl group end. In silico docking analyses identified the lipophilic interactions within the central cavity as the main driving force for binding with electrostatic interactions and H-bonds playing a minor role.

Modification of IgE binding to αS1-casein by proteolytic activity of Enterococcus faecium isolated from Iranian camel milk samples.

Milk is a perfect source of nutrients for neonates. When breast feeding cannot be done, an infant's alimentation is usually initiated to cow's milk, among the primary foods. It has been reported that about 2.5% of juveniles under the age of 3 years manifest allergic reactions to cow's milk proteins. Among the cow's milk proteins, casein fractions are considered as the strongest allergenic proteins. The proteolytic enzymes of lactic acid bacteria (LAB), during fermentation of dairy products, can break down milk proteins especially caseins and subsequently reduce the immune reactivity of allergenic proteins. In this research, raw bovine and camel milk samples were screened for cocci LAB strains and after isolation, their proteolytic activity against bovine milk caseins were evaluated by SDS-PAGE and RP-HPLC. The potential of cocci LAB strains on αS1-casein degradation and their potential to break down the principle allergenic epitopes of this protein was detected using indirect competitive ELISA. Molecular identification of the best proteolytic strain was fulfilled by 16S rDNA fragment sequencing with universal primers. The obtained results demonstrated that Enterococcus faecium isolated from raw camel milk samples was the most efficient isolate in hydrolyzing Na-caseinate and αS1-casein. Hydrolysated αS1-casein by Enterococcus faecium was also less recognized by IgE of bovine milk allergic patients' sera in comparison with native αS1-casein. It has been proposed that Enterococcus faecium could be an efficient strain in allergenicity reduction of cow's milk proteins. So it could be an excellent candidate to be potentially used in dairy industries.

Beneficial Protective Role of Endogenous Lactic Acid Bacteria Against Mycotic Contamination of Honeybee Beebread.

The purpose of this article is to reveal the role of the lactic acid bacteria (LAB) in the beebread transformation/preservation, biochemical properties of 25 honeybee endogenous LAB strains, particularly: antifungal, proteolytic, and amylolytic activities putatively expressed in the beebread environment have been studied. Seventeen fungal strains isolated from beebread samples were identified and checked for their ability to grow on simulated beebread substrate (SBS) and then used to study mycotic propagation in the presence of LAB. Fungal strains identified as Aspergillus niger (Po1), Candida sp. (BB01), and Z. rouxii (BB02) were able to grow on SBS. Their growth was partly inhibited when co-cultured with the endogenous honeybee LAB strains studied. No proteolytic or amylolytic activities of the studied LAB were detected using pollen, casein starch based media as substrates. These findings suggest that some honeybee LAB symbionts are involved in maintaining a safe microbiological state in the host honeybee colonies by inhibiting beebread mycotic contaminations, starch, and protein predigestion in beebread by LAB is less probable. Honeybee endogenous LAB use pollen as a growth substrate and in the same time restricts fungal propagation, thus showing host beneficial action preserving larval food. This study also can have an impact on development of novel methods of pollen preservation and its processing as a food ingredient.

Inhibition of Prion Propagation by 3,4-Dimethoxycinnamic Acid.

Neurodegenerative diseases are associated with accumulation of amyloid-type protein misfolding products. Prion protein (PrP) is known for its ability to aggregate into soluble oligomers that in turn associate into amyloid fibrils. Preventing the formation of these infective and neurotoxic entities represents a viable strategy to control prion diseases. Numerous attempts to find dietary compounds with anti-prion properties have been made; however, the most promising agent found so far was curcumin, which is poorly soluble and merely bioavailable. In the present work, we identify 3,4-dimethoxycinnamic acid (DMCA) which is a bioavailable coffee component as a perspective anti-prion compound. 3,4-Dimethoxycinnamic acid was found to bind potently to prion protein with a Kd of 405 nM. An in vitro study of DMCA effect on PrP oligomerization and fibrillization was undertaken using isothermal titration calorimetry (ITC), dynamic light scattering (DLS) and circular dichroism (CD) methodologies. We demonstrated that DMCA affects PrP oligomer formation reducing the oligomer content by 30-40%, and enhancing SH-SY5Y cell viability treated with prion oligomers. Molecular docking studies allowed to suggest a site where DMCA is able to bind stabilizing PrP tertiary structure. We suggest that DMCA is a perspective dietary compound for prophylaxis of neurodegenerative diseases that needs further research. Copyright © 2017 John Wiley & Sons, Ltd.

ß-Lactoglobulin: An efficient nanocarrier for advanced delivery systems.

Thanks to the progress of nanotechnology there are several agent-delivery systems that can be selected to achieve rapid and specific delivery of a wide variety of biologically active agents. Consequently, the manipulation and engineering of biopolymers has become one of the most exciting subjects for those who study delivery systems on the nanoscale. In this regard, both nanoparticle formation and a carrier role have been observed in the case of the globular milk whey protein, ß-lactoglobulin (ß-LG), setting it apart from many other proteins. To date, many efforts adopting different approaches have created ß-LG nanoparticles useful in forming delivery systems for various agents with specific targets. In this review, the potential of ß-LG to play the role of an efficient and diverse carrier protein, as well as its ability to form a well-targeted nano-scale delivery system is discussed.