Protein-based Formulations Research Articles

Textural improvement of pea protein-based high-moisture extrudates with corn zein and rice starch

High moisture extrusion allows the production of plant protein-based products, including meat analogues. Building upon our previous findings showing that zein mixed with rice starch provides the necessary textural properties to formulations, different pea protein-based formulations with varying amounts of zein and rice starch or wheat gluten (as control) were produced using high moisture extrusion and the rheological, textural, and microstructural characteristics were evaluated and associated with the secondary structure of proteins. Samples containing wheat gluten presented desirable rheological and mechanical properties in terms of texturization, which was evidenced by the generation of a layered and three-dimensional viscoelastic network. The addition of rice starch to zein significantly increased the viscoelasticity of the samples due to enhanced development of non-covalent interactions that led to higher and more stable β-sheets content and to the formation of a fibrous and layered microstructure and a 3D network nearly like those obtained with gluten. The sole replacement of pea protein by zein was not enough to develop these desired characteristics, demonstrating the importance of the non-covalent interactions between rice starch and zein for the generation of these properties. Overall, zein and rice starch improved texturization of pea protein-based gluten-free analogues made by high moisture extrusion.

Improved encapsulation efficiency and storage stability of lutein by soy protein isolate nanocarriers with thermal and trypsin treatments.

Encapsulation of bioactive compounds within protein-based nanoparticles has garnered considerable attention in the food and pharmaceutical industries because of its potential to enhance stability and delivery. Soy protein isolate (SPI) has emerged as a promising candidate, prompting the present study aiming to modify its properties through controlled thermal and trypsin treatments for improved encapsulation efficiency (EE) of lutein and its storage stability. The EE of lutein nanoparticles encapsulated using SPI trypsin hydrolysates (SPIT) with three varying degrees of hydrolysis (4.11%, 6.91% and 10.61% for SPIT1, SPIT2 and SPIT3, respectively) increased by 12.00%, 15.78% and 18.59%, respectively, compared to SPI. Additionally, the photostability of SPIT2 showed a remarkable increase of 38.21% compared to SPI. The superior encapsulation efficiency and photostability of SPIT2 was attributed to increased exposure of hydrophobic groups, excellent antioxidant activity and uniform particle stability, despite exhibiting lower binding affinity to lutein compared to SPI. Furthermore, in SPIT2, the protein structure unfolded, with minimal impact on overall secondary structure upon lutein addition. The precise application of controlled thermal and trypsin treatments to SPI has been shown to effectively produce protein nanoparticles with substantially improved encapsulation efficiency for lutein and enhanced storage stability of the encapsulated lutein. These findings underscore the potential of controlled thermal and trypsin treatments to modify protein properties effectively and offer significant opportunities for expanding the applications of protein-based formulations across diverse fields. © 2024 Society of Chemical Industry.

Influence of Nano-Particulate Impurities and β-glucans on the Stability of Protein-Based Formulations.

Pharmaceutical grade sugars manufactured under Current Good Manufacturing Practice (cGMP) and complied with International Pharmaceutical Excipients Council (IPEC) quality standards, also contain a significant amount of nano-particulate impurities (NPIs). This review will focus on the origin of NPIs, the mechanism of their interference with Dynamic light scattering (DLS) and endotoxin tests, filtration technology to effectively reduce the NPIs, methodologies for analytical quantification of NPIs, guidance for setting the limits of threshold concentration and the overall impact of NPIs on the therapeutic activity, performance, stability of biopharmaceuticals and protein-based formulations. NPIs with an average particle size of 100 to 200 nm are present in sugars and are a combination of various chemicals such as dextrans (with the presence of β-glucans), ash, inorganic metal salts, aromatic colorants, etc. These NPIs primarily originate from raw materials and cannot be removed during the sugar refinement process. While it is commonly believed that filtering the final formulation with a 0.22 μ sterilizing grade filter removes all microbes and particles, it is important to note that NPIs cannot be filtered using this standard sterile filtration technology. Exceeding the threshold limit of NPIs can have detrimental effects on formulations containing proteins, monoclonal Antibodies (mAbs), nucleic acids, and other biopharmaceuticals. NPIs and β-glucans have a critical impact on the functionality and therapeutic activity of biomolecules and if present below the threshold limit of reaction, stability and shelf-life of biologics formulation will be greatly improved and the risk of immunogenic reactions must be significantly decreased.

Current Challenges and Approaches on Delivery of Protein Based Formulations

Numerous obstacle crops up in the path of developing efficient protein-based formulations and administration in humans for their therapeutic effect. The challenges starts from protein synthesis to it’s handling procedure, stability, storage, excipients compatibility, scale up, analysis, validation, tissue targeting, in-vivo bioavailability, immunogenic reactions, etc., In the field of protein drug development, various novel methods and techniques have been evolved and continuing the research to face those challenges to bring out potential drug formulations and its delivery system to the market. It is vital to develop various strategies for protecting the integrity of proteins, increasing their availability in the body, decreasing immunological responses, transporting and target to particular tissues or cells. This current review focuses on to understand the problems, challenges encountering and the best approaches for protein based drug formulation development. It is one of roadmap to plan various research activities for continual improvement of therapeutic applications of newly developing protein drugs in this area.

Nanoparticles in Clinical Trials: Analysis of Clinical Trials, FDA Approvals and Use for COVID-19 Vaccines.

Nanoparticles are heterologous small composites that are usually between 1 and 100 nanometers in size. They are applied in many areas of medicine with one of them being drug delivery. Nanoparticles have a number of advantages as drug carriers which include reduced toxic effects, increased bioavailability, and their ability to be modified for specific tissues or cells. Due to the exciting development of nanotechnology concomitant with advances in biotechnology and medicine, the number of clinical trials devoted to nanoparticles for drug delivery is growing rapidly. Some nanoparticles, lipid-based types, in particular, played a crucial role in the developing and manufacturing of the two COVID-19 vaccines-Pfizer and Moderna-that are now being widely used. In this analysis, we provide a quantitative survey of clinical trials using nanoparticles during the period from 2002 to 2021 as well as the recent FDA-approved drugs (since 2016). A total of 486 clinical trials were identified using the clinicaltrials.gov database. The prevailing types of nanoparticles were liposomes (44%) and protein-based formulations (26%) during this period. The most commonly investigated content of the nanoparticles were paclitaxel (23%), metals (11%), doxorubicin (9%), bupivacaine and various vaccines (both were 8%). Among the FDA-approved nanoparticle drugs, polymeric (29%), liposomal (22%) and lipid-based (21%) drugs were the most common. In this analysis, we also discuss the differential development of the diverse groups of nanoparticles and their content, as well as the underlying factors behind the trends.

Matching textural properties of commercial meat and cheese products using zein as the viscoelastic agent and calcium hydroxide as the textural modifier in plant-based formulations

Plant-based proteins have the potential to replace animal proteins in food applications; however, they do not provide cohesive and viscoelastic properties to mimic such products. Matching textural properties is one of the biggest challenges in plant-based formulations. In this study, building upon our previous work, we developed a zein-pea protein-based formulation and adjusted the textural properties by varying the calcium hydroxide concentration to match the textural properties of commercial meat and cheese products. Weak frequency dependence and higher storage modulus compared to loss modulus suggested solid-like behavior in the dough-like blends, possessing a strong and elastic inner network that retained energy inside the structure rather than dissipating it. Imaging using scanning electron and confocal microscopies showed well-incorporated web-like networks of zein conferring the viscoelastic and cohesive structure. Infrared spectroscopy (FT-IR) showed that calcium hydroxide addition at various levels (1–7%) had no significant effect on the product's molecular and structural chemistry. A high amount of β-sheet structure was found in all formulations, resulting in good viscoelastic characteristics. Notably, textural properties (i.e., hardness, chewiness, gumminess) of the formulated ingredient blends could be matched to those of tested commercial products by changing the calcium hydroxide concentration (1%-tilapia fish, 3%-chicken nugget and mozzarella cheese, 5%-tuna fish and Colby cheese, 7%-burger patties). Practical outcomes are formulations of new plant-based analogues that match the textural properties of existing animal-based commercial products.

3D Printing of Textured Soft Hybrid Meat Analogues

Meat analogue is a food product mainly made of plant proteins. It is considered to be a sustainable food and has gained a lot of interest in recent years. Hybrid meat is a next generation meat analogue prepared by the co-processing of both plant and animal protein ingredients at different ratios and is considered to be nutritionally superior to the currently available plant-only meat analogues. Three-dimensional (3D) printing technology is becoming increasingly popular in food processing. Three-dimensional food printing involves the modification of food structures, which leads to the creation of soft food. Currently, there is no available research on 3D printing of meat analogues. This study was carried out to create plant and animal protein-based formulations for 3D printing of hybrid meat analogues with soft textures. Pea protein isolate (PPI) and chicken mince were selected as the main plant protein and meat sources, respectively, for 3D printing tests. Then, rheology and forward extrusion tests were carried out on these selected samples to obtain a basic understanding of their potential printability. Afterwards, extrusion-based 3D printing was conducted to print a 3D chicken nugget shape. The addition of 20% chicken mince paste to PPI based paste achieved better printability and fibre structure.

Current Stage in the Development of Klebsiella pneumoniae Vaccines.

Klebsiella pneumoniae is a bacterium capable of colonizing mucous membranes, causing serious infections. Widespread antibiotic resistance in K. pneumoniae—either through intrinsic mechanisms or via acquisition from different species, especially in hospital environments—limits the therapeutic options against this pathogen, further aggravating the disease burden. To date, there are no vaccines available against K. pneumoniae infection. Although formulations based on capsular polysaccharides have been proposed, the high variability in capsular serotypes limits vaccine coverage. Recombinant vaccines based on surface exposed bacterial antigens are a promising alternative owing to their conservation among different serotypes and accessibility to the immune system. Many vaccine candidates have been proposed, some of which have reached clinical trials. The present review summarizes the current status of K. pneumoniae vaccine development. Different strategies including whole cell vaccines, outer membrane vesicles (OMVs), ribosome, polysaccharide, lipopolysaccharide (LPS), and protein-based formulations are discussed. The contribution of antibody and cell-mediated responses is also presented. In summary, K. pneumoniae vaccines are feasible and a promising strategy to prevent infections and to reduce the antimicrobial resistance burden worldwide.

Chemical stabilization of γ-polyglutamate by chitosan and the effect of co-solvents on the stability

In protein-based formulations, conformational distortions and attractive interactions may cause insoluble and undesired aggregates. In the case of ionic peptides, including cationic or anionic, commonly electrostatic interactions are the main factors that control structure assembling. In this study, it was proposed that grafting of chitosan (CS) to γ-polyglutamic acid (γ-PGA) might exhibit much strong inhibiting effect on the formation of protein aggregates due to multiple amino groups and hydrophilic properties. To guarantee stable and safe biopharmaceutical formulation, the potency of a variety of stabilizers including sugars (glucose, sucrose), polyols (sorbitol, glycerol), surfactant (Tween 20), salting-out salt (PBS), and also different pH values have been evaluated on stabilizing or destabilizing the native state of CS-g-PGA copolymer using FTIR, CD, DLS, and SDS-PAGE. The comparable analysis revealed that the stability of CS-g-PGA was strongly dependent on pH owing to the polyelectrolyte characteristics of the polymers. Altogether these results implied that CS at optimized conditions might be an important precursor for the pharmaceutical industry and function as a new polymer for aggregation suppression and protein stabilization.

LYOPHILIZATION: AN IMPORTANT FORMULATION TECHNIQUE

Lyophilization, is an important development for medicament formulation because of its ability to safely dry heat-sensitive vaccines, antibiotics, and protein-based formulations. The lyophilization process results in long shelf lives powder formulation which can be reconstituted at the point of use. As Injectable biopharmaceuticals become a more prominent part of the overall drug market the value of lyophilized product is has increased significantly. In 1998, lyophilized pharmaceuticals accounted for 11.9% of all new Injectable or infusible drugs, but, by 2015, they made up half of all such new drug introductions (Ratti et al 2001, Bubnovich et al 2012).

Evaluation of graded levels of soy oil as a fish oil replacement in high soy protein feeds for juvenile red drum, Sciaenops ocellatus

A 9-week feeding trial was conducted with juvenile red drum, Sciaenops ocellatus, to evaluate the use of soy oil as a fish oil replacement. Three primary protein sources (fishmeal - FM, soybean meal - SBM, and soy protein concentrate - SPC) were utilized with 100% fish oil (FM, SBM, SPC), 75% fish oil (SBM, SPC), or 50% fish oil (FM, SBM, SPC) as the lipid source. Traditional growth and performance metrics (specific growth rate, feed consumption, feed conversion ratio) were tracked and tissue samples (liver, muscle, plasma, adipose, and brain) were collected for gas chromatography-based fatty acid profiling. Ten lipid metabolism related genes were analyzed for potential expression differences between dietary treatments in liver and muscle tissues and whole body and fillet tissues were sampled for proximate composition analyses. Forty- four fatty acids were measured by gas chromatography-flame ionization detector (GC-FID) and evaluated with principle component analysis and ANOVA to understand the dietary influence on lipid metabolism and health. Significant differences in growth rate were observed with the SBM 50% fish oil diet outperforming the FM 100% fish oil reference diet. All other soy protein-based diets performed statistically equivalent to both FM reference diets (100% and 50% fish oil) in regard to growth, however all soy protein-based formulations had significantly lower feed conversion ratios than the fishmeal-based references (p < .001). Gene expression differences were not significant in most cases, however often trended similarly as the observed performance. Fatty acid profiles differed as a function of oil source, with no apparent influence by protein source, with C18:2n-6 (linoleic acid) being-the primary differentiator. Overall, the six soy protein, fishmeal-free formulations performed equivalently or better than the fishmeal references with up to 50% of fish oil replaced by soybean oil.

A model-based approach for the rational design of the freeze-thawing of a protein-based formulation

Proteins are unstable molecules that may be severely injured by stresses encountered during freeze-thawing. Despite this, the selection of freeze-thaw conditions is currently empirical, and this results in reduced process control. Here we propose a mathematical model that takes into account the leading causes of protein instability during freeze-thawing, i.e. cold denaturation and surface-induced unfolding, and may guide the selection of optimal operating conditions. It is observed that a high cooling rate is beneficial for molecules that are extremely sensitive to cold denaturation, while the opposite is true when ice-induced unfolding is dominant. In all cases, a fast thawing rate is observed to be beneficial. The simulation outputs are confirmed by experimental data for myoglobin and lactate dehydrogenase, suggesting that the proposed modeling approach can reproduce the main features of protein behavior during freeze-thawing. This approach can therefore guide the selection of optimal conditions for protein-based formulations that are stored in a frozen or freeze-dried state.

GPU acceleration of Fitch’s parsimony on protein data: from Kepler to Turing

The analysis of complex biological datasets beyond DNA scenarios is gaining increasing interest in current bioinformatics. Particularly, protein sequence data introduce additional complexity layers that impose new challenges from a computational perspective. This work is aimed at investigating GPU solutions to address these issues in a representative algorithm from the phylogenetics field: Fitch’s parsimony. GPU strategies are adopted in accordance with the protein-based formulation of the problem, defining an optimized kernel that takes advantage of data parallelism at the calculations associated with different amino acids. In order to understand the relationship between problem sizes and GPU capabilities, an extensive evaluation on a wide range of GPUs is conducted, covering all the recent NVIDIA GPU architectures—from Kepler to Turing. Experimental results on five real-world datasets point out the benefits that imply the exploitation of state-of-the-art GPUs, representing a fitting approach to address the increasing hardness of protein sequence datasets.

Influence of protein source on the morphology, physicochemical and flow properties of protein-based emulsion particles to be used as texture modulators

The present study compared the influence of heating (95 °C for 90 s) formulations based on globular proteins with similar isoelectric points (whey, pea, potato and soy), with the same nutrient composition (3 wt% protein; pH 6.6; 0–0.95 wt% NaCl; 0–0.10 wt% CaCl2; 0–5 wt% high oleic sunflower oil) and processed using the same conditions (homogenization and shearing post heat treatment), on the morphology, physicochemical and flow properties of the resulting aggregated systems. Some trends in the aggregation behavior among the different protein-based formulations were identified, but the extent of the modifications promoted by heat treatment was protein source-dependent. Heat treatment of the oil-free formulations in the presence of added salts resulted in extensive protein aggregation, as inferred from the considerable decrease in protein solubility and increase in particle size. Interestingly, addition of oil to the various salts-containing formulations led to a further increase in the aggregation yield and a decrease in particle size. The morphology of the aggregates differed considerably depending on the protein source used, with oil addition leading to a loss of protein network. Furthermore, addition of oil to plant protein-based formulations promoted a marked increase in viscosity upon heat treatment, whereas the opposite phenomenon was observed for whey protein-based formulations. The differences observed among the various protein-based aggregated systems in terms of morphology, physicochemical and flow properties may be ascribed to differences in their sensitivity towards added salts and emulsifying ability, as observed before heat treatment, as well as in their denaturation temperature, given the relatively short heating time used in this study. Therefore, formulation and processing need to be adapted to the protein used if aggregated systems with similar properties want to be obtained starting from different protein sources under intermediate degrees of electrostatic repulsion.

Dehydration of protein lactoferrin-glycomacropeptide nanohydrogels

Protein based nanohydrogels have recently gained interest due their high ability to carry and deliver active compounds. However, one of the limitations of using protein-based systems is their high sensitivity to physicochemical stresses during processing and storage. Protein-based formulations are commonly prepared as solid dosage forms once their stability can be achieved in the solid rather than in the liquid state. One of the methods to improve the stability of protein-based systems is drying, being freeze and spray drying the most popular methods. Nanohydrogels composed of lactoferrin and glycomacropeptide were developed by thermal gelation and dried using a freeze-dryer (CHRIST - Alpha 1–4 LD plus, Germany) and a nano spray dryer B-90 (BÜCHI Labortechnik AG, Switzerland). For nano spray-drying different experimental conditions were tested such as: the inlet temperature (80, 100 and 120 °C) and the hole size of the vibrating membrane in the spray cap (4 μm and 7 μm), while for freeze drying the nanohydrogels where maintained at −40 °C for a drying period of about 24 h. The physicochemical properties of the resulting dried nanohydrogels were compared through different techniques: differential scanning calorimetry (DSC), thermogravimetry (TGA), X-ray diffraction (XRD) and scanning electron microscopy (SEM). DSC shows an endothermic peak ranging from 82 to 90 °C, indicating that during the drying methodologies the second denaturation peak of lactoferrin (around 90 °C) is maintained. No significant differences (p < 0.05) for denaturation peak temperatures were observed when different inlet temperatures or spray cap size were applied during the nano spray-drying conditions. When compared with freeze-drying it is possible to observe that a denaturation peak around the same range of temperatures was detected, also indicating that this drying process did not affect the denaturation of the protein. XRD results of nanohydrogels dried by freeze-drying revealed that crystallinity is significantly higher when compared with spray-dried nanohydrogels, explaining the lower thermal degradation observed in TGA for freeze-dried nanohydrogels. Nanohydrogels morphology after the drying process was evaluated by SEM. Nanohydrogels obtained by nano spray dryer B-90 allowed identifying the presence of particles with spherical shape, while nanohydrogels obtained by freeze drying had a sponge-like appearance. This work shows the influence of the drying process of protein-based nanohydrogels in their physical-chemical properties, thus pointing at which will be the preferred technology to be used in a future industrial application.

Antioxidant activity and characterization of whey protein-based beverages: Effect of shelf life and gastrointestinal transit on bioactivity

Abstract Whey proteins can exhibit antioxidant activity. The objectives of this study were to formulate model whey based beverages with well-established antioxidants (plant polyphenols, vitamins and astaxanthin) to investigate (1) the antioxidant shelf life over a 24-week period and (2) the antioxidant activity after upper gastrointestinal transit. Pilot scale processing (pasteurization, ultra-high temperature or spray drying) was used to prepare beverages which were representative of current product formats. In vitro gastrointestinal digestion of test samples was performed using the standardised INFOGEST method and antioxidant activity of samples was determined using ABTS, FRAP and ORAC. Results from the antioxidant shelf life study provided evidence that powder products functionality was preserved. Whey beverages (pasteurised or spray dried) increased or maintained antioxidant activity during gastrointestinal transit. Combination of whey with additional antioxidant ingredients increased the bioactivity of formulated products; however, this greater bioactivity was altered after gastrointestinal transit, depending on processing type and antioxidant methodology. Industrial relevance Whey protein-based antioxidant beverages could benefit the elderly consumer to meet their increased protein requirements and boost their antioxidant status. Consumer's acceptance for whey protein-based beverages often improves with clear formulations. This work generated whey protein-based UHT beverages with greater stability and clarity than pasteurised formulations. A novel combination of plant and marine antioxidants increased antioxidant activity of whey protein-based formulations. Furthermore, to suit export markets this work generated spray dried whey protein formulations that did not alter antioxidant potential.

Effect of Arginine on the Aggregation of Protein in Freeze-Dried Formulations Containing Sugars and Polyol: II. BSA Reconstitution and Aggregation

The current paper continues our study on the ability of L-arginine to prevent/reduce the aggregation of proteins that results from the various stresses during the lyophilisation and/or storage of lyophilized protein-based products. The first part of our study, i.e. formulation development, was devoted to the rational design and optimization of an L-arginine containing lyophilized formulation which can resist the natural tendency of L-arginine to absorb atmosphere moisture. Mannitol and trehalose were chosen among other excipients to be included in the protein-based formulation, as mannitol in a combination with L-arginine has been shown to reduce moisture sorption while trehalose provides a degree of lyoprotection. In the present study, a number of formulations, which comprised bovine serum albumin (BSA) with and without L-arginine, and with five different ratios of trehalose-to-mannitol (from 30:70 to 80:20) were lyophilised and assessed. The internal structures and the moisture sorption/retention of the lyophilized formulations were characterised. To study the effect of L-arginine on BSA solid-phase stability, the lyophilized powder was exposed to accelerated storage conditions (high moisture (75% RH) and temperature (22 or 45°C)) for up to 24h. The lyophilized BSA formulations were then reconstituted and solution-state protein aggregation assessed by turbidimetry at 360nm and fluorescence spectroscopy using the thioflavin T assay. It was demonstrated that L-arginine can be used in protein-based freeze-dried formulations to significantly reduce the aggregation of protein during the manufacturing, storage and subsequent reconstitution. The results also revealed the importance of a sufficient amount of mannitol in the arginine-containing formulations.

Protein based devices for oral tissue repair and regeneration

In the last decades, a goal of tissue engineering has been devoted to the design of devices with multiple micro- or nano-structures and loaded with bioactive molecules, to mimic the extracellular matrix (ECM) so generating a conducive microenvironment for new tissue replacement/regeneration. The ECM, naturally, is composed of fibrous proteins which provide structural support for tissues, mainly regulating cells behavior in terms of proliferation, growth, survival, shape, migration and differentiation by cell-matrix interactions. Several studies have been just investigated the fabrication of different platforms for the regeneration of teeth, oral mucosa, salivary glands, bone, and periodontium. In this context, many proteins—from a natural or biological source—have been used as instructive substances to in vitro guide tissue organization and functions. In particular, new advances in the definition of protein-based formulations currently represent a great challenge to promote a more effective regeneration of dental tissues to be transplanted into patients to replace damaged, diseased or missing tissues. Hence, the purpose of this review is to discuss the use of protein-based systems for the regeneration of oral tissues.

Computational methods in preformulation study for pharmaceutical solid dosage forms of therapeutic proteins

AbstractDesign and delivery of protein-based biopharmaceuticals needs detailed planning and strict monitoring of intermediate processing steps, storage conditions and container-closure system to ensure a stable, elegant and biopharmaceutically acceptable dosage form. Selection of manufacturing process variables and conditions along with packaging specifications can be achieved through properly designed preformulation study protocol for the formulation. Thermodynamic stability and biological activity of therapeutic proteins depend on folding–unfolding and three-dimensional packing dynamics of amino acid network in the protein molecule. Lack of favourable environment may cause protein aggregation with loss in activity and even fatal immunological reaction. Although lyophilization can enhance the stability of protein-based formulations in the solid state, it can induce protein unfolding leading to thermodynamic instability. Formulation stabilizers such as preservatives can also result in aggregation of therapeutic proteins. Modern instrumental techniques in conjunction with computational tools enable rapid and accurate prediction of amino acid sequence, thermodynamic parameters associated with protein folding and detection of aggregation “hot-spots.” Globular proteins pose a challenge during investigations on their aggregation propensity. Biobetter therapeutic monoclonal antibodies with enhanced stability, solubility and reduced immunogenic potential can be designed through mutation of aggregation-prone zones. The objective of the present review article is to focus on the various analytical methods and computational approaches used in the study of thermodynamic stability and aggregation tendency of therapeutic proteins, with an aim to develop optimal and marketable formulation. Knowledge of protein dynamics through application of computational tools will provide the essential inputs and relevant information for successful and meaningful completion of preformulation studies on solid dosage forms of therapeutic proteins.

Surface modulation of polymeric nanocarriers enhances the stability and delivery of proteins and small molecules.

We aimed to enhance the stability and therapeutic efficiency of protein-based therapeutic formulations. Proteins were immobilized on the surface of nanoparticles (NPs) to improve both protein stability and protein function, especially enzymatic activity. The modularity of the platform was demonstrated by coating proteins of varied molecular weights and functionalities on the surface of poly(lactic-co-glycolic acid)-based NPs. Coating proteins to the particle surface greatly enhanced the stability of the NPs, preventing particle aggregation and improving enzymatic potency, including in vivo. Specifically, coating of collagenase I to the particle surface greatly improved the ability of the enzyme to degrade tumor collagen relative to free enzyme, thereby increasing the penetration of adjuvant chemotherapy (doxorubicin). Additionally, the protein coating reduced the rate of doxorubicin release, enabling sustained release of the small-molecule payload. The straightforward procedure described herein permits the formulation of modular NPs that can combine and sustain the benefits of small molecules and biologics.