Influence of processing factors on the stability of model mayonnaise with whole egg during long-term storage

Mayonnaise-like oil-in-water emulsions with different stabilities—evaluated from the degree of macroscopic defects, e.g., syneresis—were prepared by different formulations and processing conditions (egg yolk weight, homogenizer speed, and vegetable oil temperature). Emulsions prepared with lower egg yolk content were destabilized for shorter periods. The long-term stability of emulsions was weakly related to initial properties, e.g., oil droplet distribution and protein coverage at the interface. Protein aggregation between oil droplets was observed and would be responsible for the instability of emulsions exhibited by the appearance defects. SDS-PAGE results for adsorbed and unadsorbed proteins at the O/W interface suggested that predominant constituents adsorbed onto the interface were egg white proteins as compared with egg yolk components when the amount of added egg yolk was low. In present condition, egg white proteins adsorbed at the O/W interface could be a bridge of neighboring oil droplets thereby causing flocculation in emulsions.     

Physical properties of emulsion-based hydroxypropyl methylcellulose films: Effect of their microstructure

The initial characteristics of emulsions and the rearrangement of the oil droplets in the film matrix during film drying, which defines its microstructure, has an important role in the physical properties of the emulsion-based films. The objective of this work was to study the effect of the microstructure (two droplet size distributions) and stability (with or without surfactant) of HPMC oil-in-water emulsions over physical properties of HPMC emulsion-based edible films. HPMC was used to prepare sunflower oil-in-water emulsions containing 0.3 or 1.0% (w/w) of oil with or without SDS, as surfactant, using an ultrasonic homogenizer. Microstructure, rheological properties and stability of emulsions (creaming) were measured. In addition, microstructure, coalescence of oil droplets, surface free energy, optical and mechanical properties and water vapor transfer of HPMC films were evaluated. Image analysis did not show differences among droplet size distributions of emulsions prepared at different oil contents; however, by using SDS the droplet size distributions were shifted to lower values. Volume mean diameters were 3.79 and 3.77μm for emulsions containing 0.3 and 1.0% without surfactant, respectively, and 2.72 and 2.71μm for emulsions with SDS. Emulsions formulated with 1.0% of oil presented higher stability, with almost no change during 5 and 3 days of storage, for emulsions with and without SDS, respectively. Internal and surface microstructure of emulsion-based films was influenced by the degree of coalescence and creaming of the oil droplets. No effect of microstructure over the surface free energy of films was found. The incorporation of oil impaired the optical properties of films due to light scattering of light. Addition of oil and SDS decreased the stress at break of the emulsion-based films. The replace of HPMC by oil and SDS produce a lower "amount" of network structure in the films, leading to a weakening of their structure. The oil content and SDS addition had an effect over the microstructure and physical properties of HPMC-based emulsions which lead to different microstructures during film formation. The way that oil droplets were structured into the film had an enormous influence over the physical properties of HPMC films.     

Water-in-silicone oil emulsion stabilizing surfactants formed from native albumin and alpha,omega-triethoxysilylpropyl-polydimethylsiloxane

Contact with hydrophobic silicones frequently leads to protein denaturation. However, it is demonstrated that albumin in water-in-silicone oil emulsions retains its native structure in the presence of a functional, triethoxysilyl-terminated silicone polymer, TES-PDMS. Both HSA and TES-PDMS were essential for the formation of stable water-in-silicone oil emulsions: attempts to generate stable emulsions using independently either the protein or the functionalized silicone as a surfactant failed. Confocal microscopy indicated that the human serum albumin (HSA) preferentially adsorbed at the oil/water interface, even in the presence of another protein (glucose oxidase). A variety of experiments demonstrated that the hydrolysis of the Si-OEt groups on the functional silicone occurred only to a limited extent, consistent with the absence of a covalent linkage between the silicone and protein, or of cross-linked silicones at the interface. The fluorescence spectra of HSA extracted from the emulsions, front-faced fluorescence experiments on the HSA/silicone emulsion itself, and HSA/salicylate binding studies all demonstrated that the stability of the water/oil interface decreased as the protein began to unfold: unfolding of the protein in the emulsion was slower than in aqueous solution. The experimental evidence indicated that the interaction between HSA and TES-PDMS is not associated with either homomolecular (HSA/HSA; TES-PDMS/TES-PDMS) interactions or with covalent linkage between two the polymers. Rather, the data is consistent with the direct binding of unhydrolyzed Si(OEt) 3 groups to native HSA. The nature of these interactions is discussed.     

Whey Protein/Polysaccharide-Stabilized Emulsions: Effect of Polymer Type and pH on Release and Topical Delivery of Salicylic Acid

Emulsions are widely used as topical formulations in the pharmaceutical and cosmetic industries. They are thermodynamically unstable and require emulsifiers for stabilization. Studies have indicated that emulsifiers could affect topical delivery of actives, and this study was therefore designed to investigate the effects of different polymers, applied as emulsifiers, as well as the effects of pH on the release and topical delivery of the active. O/w emulsions were prepared by the layer-by-layer technique, with whey protein forming the first layer around the oil droplets, while either chitosan or carrageenan was subsequently adsorbed to the protein at the interface. Additionally, the emulsions were prepared at three different pH values to introduce different charges to the polymers. The active ingredient, salicylic acid, was incorporated into the oil phase of the emulsions. Physical characterization of the resulting formulations, i.e., droplet size, zeta potential, stability, and turbidity in the water phase, was performed. Release studies were conducted, after which skin absorption studies were performed on the five most stable emulsions, by using Franz type diffusion cells and utilizing human, abdominal skin membranes. It was found that an increase in emulsion droplet charge could negatively affect the release of salicylic acid from these formulations. Contrary, positively charged emulsion droplets were found to enhance dermal and transdermal delivery of salicylic acid from emulsions. It was hypothesized that electrostatic complex formation between the emulsifier and salicylic acid could affect its release, whereas electrostatic interaction between the emulsion droplets and skin could influence dermal/transdermal delivery of the active.     

Emulsions of oil from Adenanthera pavonina L. seeds and their protective effect

In our previous study, we developed very stable formulations of submicron oil-in-water emulsions from Adenanthera pavonina L. (family Leguminosae, subfamily Mimosoideae) seed oil, stabilised with soybean lecithin (SPC). Continuing our research, we introduced an additional co-emulsifier, Tween 80, to those formulations in order to decrease the size of the emulsion particles and improve their stability. Formulations with a mean particle size ranging from 43.6 to 306.5 nm and a negative surface charge from −45.3 to −28.5 mV were obtained. Our stability experiments also revealed that most of the tested formulations had a very good degree of stability over a 3-month storage period, both at 4°C and at room temperature. Since many intravenous injectable drugs exhibit lytic activity against erythrocytes, we examined this activity for the emulsion form of cardol, a natural compound with already proven hemolytic properties. The incorporation of this agent into the emulsion caused an evident decrease in hemolytic activity (97–99%). This highly protective effect, observed against sheep erythrocytes, was independent of both the composition and the particle size of the emulsions used. Our studies suggest that nonionic surfactant/phospholipid-based emulsions containing this edible oil of A. pavonina L. may be useful as an alternative formulation matrix for pharmaceutical, nutritional or cosmetic applications of otherwise membrane-acting components.     

Delayed hypersensitivity and granulomatous response after immunization with protein antigens associated with a mycobacterial glycolipid and oil droplets.

A myocardial glycolipid (P3) mixed with protein antigens in oil-in-water emulsion induced lasting delayed hypersensitivity (DH) and granulomatous inflammation after intradermal injection into guinea pigs. This did not occur when P3 and bovine serum albumin (BSA) were given in Freund's incomplete adjuvant. The oil-in-water emulsions consisted of microscopic oil droplets suspended in aqueous medium. By separating oil and aqueous phases from BSA + P3 emulsion it was shown that antigen retained with oil droplets led to DH and granuloma formation. The association of antigen with oil droplets was P3 dependent and was quantitated with 125I-labeled BSA. The same phenomenon occurred with 125I-labeled rabbit gamma-globulin (RGG) + P3 emulsion. Fluorescein-conjugated RGG was observed in a particulate state within or on oil droplets in emulsion containing P3. These physical characteristics of antigen + P3 emulsion appeared to be important for immunogenicity.     

Deep learning image analysis models pretrained on daily objects are useful for the preliminary characterization of particulate pharmaceutical samples

Visible and subvisible particles are a quality attribute in sterile pharmaceutical samples. A common method for characterizing and quantifying pharmaceutical samples containing particulates is imaging many individual particles using high-throughput instrumentation and analyzing the populations data. The analysis includes conventional metrics such as the particle size distribution but can be more sophisticated by interpreting other visual/morphological features. To avoid the hurdles of building new image analysis models capable of extracting such relevant features from scratch, we propose using well-established pretrained deep learning image analysis models such as EfficientNet. We demonstrate that such models are useful as a prescreening tool for high-level characterization of biopharmaceutical particle image data. We show that although these models are originally trained for completely different tasks (such as the classification of daily objects in the ImageNet database), the visual feature vectors extracted by such models can be useful for studying different types of subvisible particles. This applicability is illustrated through multiple case studies: (i) particle risk assessment in prefilled syringe formulations containing different particle types such as silicone oil, (ii) method comparability with the example of accelerated forced degradation, and (iii) excipient influence on particle morphology with the example of Polysorbate 80 (PS80). As examples of agnostic applicability of pretrained models, we also elucidate the application to two high-throughput microscopy methods: microflow and background membrane imaging. We show that different particle populations with different morphological and visual features can be identified in different samples by leveraging out-of-the-box pretrained models to analyze images from each sample.     

Impact of Heat and Laccase on the pH and Freeze-Thaw Stability of Oil-in-Water Emulsions Stabilized by Adsorbed Biopolymer Nanoparticles

The enzymatic cross-linking of adsorbed biopolymer nanoparticles formed between whey protein isolate (WPI) and sugar beet pectin using the complex coacervation method was investigated. A sequential electrostatic depositioning process was used to prepare emulsions containing oil droplets stabilized by WPI – nanoparticle – membranes. Firstly, a finely dispersed primary emulsion (10 % w/w miglyol oil, 1 % w/w WPI, 10 mM acetate buffer at pH 4) was produced using a high-pressure homogenizer. Secondly, a series of biopolymer particles were formed by mixing WPI (0.5 % w/w) and pectin (0.25 % w/w) solutions with subsequent heating above the thermal denaturation temperature (85 °C, 20 min) to prepare dispersions containing particles in the submicron range. Thirdly, nanoparticle-covered emulsions were formed by diluting the primary emulsion into coacervate solutions (0–0.675 % w/w) to coat the droplets. Oil droplets of stable emulsions with different interfacial membrane compositions were subjected to enzymatic cross-linking. We used cross-linked multilayered emulsions as a comparison. The pH stability of primary emulsions, biopolymer complexes and nanoparticle-coated base emulsions, as well as multilayered emulsions, was determined before and after enzyme addition. Freeze-thaw stability (?9 °C for 22 h, 25 °C for 2 h) of nanoparticle-coated emulsions was not affected by laccase. Results indicated that cross-linking occurred exclusively in the multilamellar layers and not between adsorbed biopolymer nanoparticles. Results suggest that the accessibility of distinct structures may play a key role for biopolymer-cross-linking enzymes.     

Emulsification Capacity of Microgels Assembled from β-Lactoglobulin and Pectin

Microgels formed from beta-lactoglobulin were used to prepare oil-in-water emulsions in order to examine their emulsifying capacity. Corn oil emulsions prepared with microgels of pure beta-lactoglobulin at pH 5.8 were initially stable, but a fraction of the droplets quickly flocculated to form a creamed layer that could not be dispersed by shear, which was attributed to hydrophobic attractions between the microgels on adjoining droplets. Emulsions prepared from microgels of beta-lactoglobulin and pectin at pH 4.75 possessed greater droplet sizes at lower concentrations, yet all emulsions were relatively stable to irreversible flocculation. Increased stability of emulsions stabilized by BP-gels was attributed to the presence of pectin on the surface of microgels, which increased repulsions between adjoining droplets. Stable corn oil emulsions were still prepared from microgels that were previously dialyzed to remove non-aggregated protein, which verified that the microgels were responsible for stabilizing emulsion droplets. Equilibrium surface pressure of corn oil droplets was similar between microgels and the unheated beta-lactoglobulin and pectin, yet the dynamic surface pressure was reduced at intermediate times and indicated a slow relaxation and deformation of the microgels at the interface. Microgels formed with pectin stabilized emulsions containing 90 % limonene for up to 5 days of room temperature storage, demonstrating the capacity of such protein microgels to stabilize flavor oil emulsions.     

Evaluation of Electrostatic Interaction between Lysolecithin and Chitosan in Two-Layer Tuna Oil Emulsions by Nuclear Magnetic Resonance (NMR) Spectroscopy

The main objective of this work was to investigate the electrostatic interaction between lysolecithin and chitosan in two-layer tuna oil-in-water emulsions using nuclear magnetic resonance (NMR) spectroscopy. The influence of chitosan concentration on the stability and properties of these emulsions was also evaluated. The 5 wt% tuna oil one-layer emulsion (lysolecithin-stabilized oil droplets without chitosan) and two-layer emulsions (lysolecithin-chitosan stabilized oil droplets) containing 5 wt% tuna oil, 1 wt% lysolecithin and various chitosan concentrations (0.025–0.40 wt%) were prepared. The one-dimensional (1D) ³¹P and ¹H NMR spectra of emulsions were then recorded at 25 °C. The results showed that addition of chitosan affected the stability and properties of lysolecithin-stabilized one-layer emulsions. The ³¹P NMR peak of the choline head group on lysolecithin molecules disappeared when chitosan was added at concentrations above neutralization concentration (> 0.05 wt%). The ¹H NMR peak intensity monitoring free amino groups (?NH ₃ ⁺) of chitosan showed a strong positive linear relationship to the chitosan concentration with a high correlation coefficient (R² ≈ 0.99). This ¹H NMR peak in emulsions could not be detected for chitosan in emulsions lower than saturation concentration (< 0.15 wt%). These phenomena indicate an electrostatic interaction between lysolecithin and chitosan at droplet surface in emulsion and were consistent with the results from zeta-potential measurements. The T ₂* relaxation time of the choline head group (N-(CH ₃)₃) signal of lysolecithin also confirmed that lysolecithin-chitosan electrostatic interaction occurs at the surface of oil droplets in two-layer emulsions. The results suggest that NMR spectroscopy can be used as an alternative method for monitoring the electrostatic interaction between surfactant and oppositely charged electrolytes or biopolymers in two-layer emulsions.     

Factors Affecting Foamed Emulsions Prepared with an Extract from <Emphasis Type="Italic">Quillaja saponaria</Emphasis> Molina: Oil Droplet Size,pH and Presence of Beta-Lactoglobulin

Oil is well-known to act as antifoam and to destabilize foam lamellae by bridging between two adjacent foam bubbles. It was hypothesized that an optimal oil droplet size exists with respect to the stability of a foamed emulsions, where the oil droplets are sufficiently small to postpone bridging and the amount of free surfactant is sufficient to stabilize the oil/water-interface and the air/water-interface. Emulsions with 0.3% Quillaja saponin and a median oil drop-let size between 0.2 and 2.0 μm were prepared under varying homogenization conditions and characterized in a dynamic foam analyzer. Results confirmed the above mentioned hypothesis. Stability of the foamed emulsions considerably increased with increasing pH, which was attributed to electrostatic repulsion between oil droplets and the effect on the balance between disjoining pressure and capillary pressure. In a binary system containing proteins and saponins, stability of foamed emulsions can be further increased when emulsifiers are added sequentially. When the emulsion is stabilized by β-LG and QS is added after emulsification stability of the foamed emulsion is distinctly higher compared to systems, where QS and β-LG are added prior to emulsification. Future studies should deepen our understanding of these complex dispersed systems by investigating the molecular interactions including other proteins and additional food constituents.     

Effect of Polyglycerol Esters of Fatty Acids on the Physicochemical Properties and Stability of β-Carotene Emulsions during Digestion in Simulated Gastric Fluid

The effects of six different polyglycerol esters of fatty acids (PGEs) and two different particle sizes produced using various processing parameters on the physicochemical properties and stability of the β-carotene emulsions during digestion in simulated gastric fluid (SGF) were investigated. β-Carotene emulsions were prepared by high-pressure homogenization using β-carotene (0.1% w/w) in soybean oil as the oil phase and 1% (w/w) PGE in Milli-Q water as the water phase. The particle size of β-carotene emulsions was measured by a laser diffraction technique, and the stability of emulsions was interpreted in terms of the increase in particle size and span value of emulsion droplets and the retention of β-carotene during digestion in SGF. The average particle size ranges of emulsions were 0.17 to 0.27 μm for fine emulsions and 1.16 to 1.59 μm for coarse emulsions. In the prepared β-carotene emulsions, the particle size decreased with increasing polymerization of the glycerol in PGEs, and the higher polymerization of the glycerol also increased the stability of emulsions during digestion in SGF. Although the β-carotene content in the emulsions significantly decreased with increasing digestion period, loss of β-carotene was more severe in unstable emulsions than in stable emulsions, suggesting that the particles incorporated into droplets could provide some protective barrier for decreasing the β-carotene degradation. Therefore, β-carotene emulsions stabilized by PGEs with high polymerization of the glycerol may be useful for further applications in food and drug formulations. Decaglycerol monooleate (MO750) was demonstrated to be the most effective emulsifier in stabilizing β-carotene emulsions in this study.     

Real-Time Determination of Structural Changes of Sodium Caseinate-Stabilized Emulsions Containing Pectin Using High Resolution Ultrasonic Spectroscopy

The interactions that lead to structure transitions in oil-in-water emulsions were investigated using high-resolution ultrasonic spectroscopy. High methoxyl pectin (HMP) was added to the emulsions at various concentrations and the dynamics of aggregation induced by changes in pH were observed. Two independent ultrasonic parameters, velocity and attenuation, were measured as a function of time or pH. At pH 6.8, both velocity and attenuation of sound changed as a function of HMP concentration. During acidification, caused by the addition of glucono-δ-lactone, there were small changes in the overall ultrasonic velocity, but it was possible to relate these changes to the structural changes in the emulsion. The values of ultrasonic attenuation decreased at high pH with increasing amount of HMP, indicating changes in the flocculation state of the oil droplets caused by depletion forces. During acidification at pH 5.4, emulsions containing HMP showed a steep increase in the ultrasonic attenuation, and this pH corresponds to the pH of association of HMP with the casein-covered oil droplets. The adsorption of HMP onto the interface causes a rearrangement of the oil droplets, and the emulsions containing sufficient amounts of HMP no longer gel at acid pH. This is well described by the ultrasonic attenuation changes in the various emulsions. This research demonstrated for the first time that ultrasonic spectroscopy can be employed for in situ monitoring and analysis of acid-induced destabilization of food emulsions.     

Particle-by-particle quantification of protein adsorption onto poly(ethylene glycol) grafted surfaces

The use and advantage of flow cytometry as a particle-by-particle, low sampling volume, high-throughput screening technique for quantitatively examining the non-specific adsorption of proteins onto surfaces is presented. The adsorption of three proteins: bovine serum albumin (BSA), immunoglobulin gamma (IgG) and protein G, incubated at room temperature for 2 h onto organosilica particles modified with poly(ethylene glycol) (PEG) of increasing MW (2000, 3400, 6000, 10,000 and 20,000 g mol(-1)) and grafted amounts (0.14-1.4 mg m(-2)) was investigated as a model system. Each protein exhibited Langmuir-like, high affinity monolayer limited adsorption on unmodified particles with the proteins reaching surface saturation at 1.8, 4.0 and 2.5 mg m(-2) for BSA, IgG and protein G, respectively. Protein adsorption on PEG-modified surfaces was found to decrease with increasing amounts of grafted polymer. PEG grafting amounts >0.6 mg m(-2) effectively prevented the adsorption of the larger two proteins (BSA and IgG) while a PEG grafting amount >1.3 mg m(-2) was required to prevent the adsorption of the smaller protein G.     

Influence of formulation additives on the desiccation tolerance and storage stability of blastospores of the entomopathogenic fungus Paecilomyces fumosoroseus (Deuteromycotina: Hyphomycetes)

Blastospores of the entomopathogenic fungus Paecilomyces fumosoroseus were formulated with 10% lactose/1% bovine serum albumin (BSA) or various compositions of Fantesk™, a starch-oil composite prepared by jet-cooking an aqueous dispersion of starch and oil. Storage stability studies with wet blastospore formulations showed that maximum blastospore survival was achieved during low-temperature storage at -20°C with lactose/BSA formulations or starch-oil formulations supplemented with sucrose, zein protein, and whole milk. Under conditions of wet storage at -20°C, the addition of whole milk to starch-oil formulations significantly improved blastospore stability while the addition of sucrose or zein protein had no effect. In freeze-drying studies, no significant differences were seen in blastospore desiccation tolerance or in stability during storage at either 4 or -20°C when blastospores of P. fumosoroseus were formulated with lactose/BSA or starch-oil formulations with sucrose, zein protein, and whole milk. Freeze-dried blastospore formulations stored at 4°C showed no loss in blastospore viability after 3 months storage and blastospore formulations stored at -20°C showed no loss in viability during the entire 12-month study. For freeze-dried, starch-oil formulations, sucrose was shown to improve blastospore survival during the freeze-drying process. The addition of whole milk to starch-oil formulations significantly improved the stability of freeze-dried blastospores stored at 4°C. Compared to unformulated blastospore suspensions that showed blastospore settling after 30 min, suspensions of blastospores formulated with lactose/BSA or starch-oil composites remained stable for up to 2 h after mixing.     

Optimization of Nanoemulsion Fabrication Using Microfluidization: Role of Surfactant Concentration on Formation and Stability

Nanoemulsions have some important potential advantages over conventional emulsions for certain commercial applications due to their optical clarity, high physical stability, and ability to increase the bioavailability of lipophilic bioactives. In this study, the factors influencing droplet size and stability in nanoemulsions fabricated from a hydrocarbon oil and an anionic surfactant were examined. Octadecane oil-in-water nanoemulsions were produced by a high pressure homogenizer (microfluidizer) using sodium dodecyl sulfate (SDS) as a model anionic surfactant. The influence of homogenization pressure, number of passes, and surfactant concentration was examined. The droplet size decreased with increasing homogenization pressure, number of passes, and surfactant concentration. Nanoemulsions with low turbidity and small droplet diameters (≈62 nm) could be produced under optimized conditions. Interestingly, nanoemulsions containing relatively high surfactant levels were highly susceptible to creaming when they were only passed through the homogenizer a few times, which was attributed to depletion flocculation. These results show the importance of optimizing surfactant levels to produce small droplets that are also stable to creaming.     

流式细胞术的发展及在植物研究中的应用

流式细胞术是通过对液流中各种荧光标记的颗粒进行多参数快速高效的定性或定量测定的方法,在科学研究的多个领域发挥重要作用。然而,由于植物组织及细胞壁和次生代谢产物等细胞的特殊成分和结构,限制了其在植物研究领域的应用。本文在介绍流式细胞仪发展和组成分类的基础上,着重讨论了流式细胞术在植物领域的应用、研究进展及应用限制,进而展望该研究领域的发展趋势,为拓宽植物流式细胞术的潜在应用范围提供新的思考方向。     

Antifreeze Emulsions Produced from Linoleic Acid and Water Using Enzymatically Modified Gelatin

An enzymatically modified gelatin with covalently attached leucine dodecyl ester, referred to as EMG-12, was used as a surfactant to prepare emulsions with different properties by changing the surfactant concentration, oil volume fraction, and pH in the water phase. The emulsions generally resisted the freezing of their constituent bulk water at approximately ?10°C, but similar emulsions produced with soy protein isolate, casein, or Tween-80 as control agents were less resistant. The freezing (or unfreezing) of the bulk water in these emulsions depended on the kind of agent used, not on the emulsion properties such as average area of the oil/water interface, stability against coalescence, and stability against creaming. The emulsion produced with EMG-12, like that produced with polyglycerol stearate, tended to maintain its unfrozen state even in the presence of silver iodide crystals added as heterogeneous ice-nuclei. The significance of producing such an antifreeze emulsion is discussed from the standpoint of cryopreservation of cold-sensitive food and biological systems.     

Influence of Ionic Surfactants on the Microstructure of Heat-Set β-Lactoglobulin-Stabilized Emulsion Gels

Using confocal microscopy, we studied the effect of heating (up to 85°C) on the microstructure of β-lactoglobulin-stabilized emulsions (20 vol% oil, pH 6.8) containing excess protein (total protein content 13.2%). Two different fluorescent dyes were used to separately visualize the oil droplets and the protein. In overlay micrographs, their location with respect to each other could then be determined. In the presence of a low salt concentration, flocculation of the emulsion without surfactant was inhibited, by a mechanism analogous to the “salting-in” of aqueous protein solutions. Addition of the anionic surfactant sodium dodecyl sulfate (SDS) caused weak flocculation, probably as a result of the formation of protein−SDS complexes. The final heat-set emulsion contained distinct pores for a surfactant/protein ratio of R = 1, but no pores for R = 2. Addition of the cationic surfactant cetyl trimethyl ammonium bromide (CTAB) caused strong aggregation, as indicated by microscopic observation of the concentrated emulsion and light scattering of the diluted emulsion. For R = 1 with CTAB, there were aggregates consisting of oil droplets and excess protein. At R = 2, almost all the excess protein was aggregated into separate protein flakes. In the final emulsion gels containing CTAB, the protein was more spread out. Differing structural behavior with anionic and cationic surfactants has been interpreted in terms of different protein−surfactant interactions in aqueous solution and at the oil−water interface, both before and after protein denaturation.     

Development of Biotechnology Products in Pre-filled Syringes: Technical Considerations and Approaches

A monoclonal antibody (mAb) product development case study is presented to address some of the issues faced during developing a pre-filled syringe (PFS) product for a biotherapeutic. In particular, issues involving incompatibility with silicone oil and a stability-based approach for selection of PFS barrel and tip cap components have been discussed. Silicone spiking studies followed by exposure to agitation stress or accelerated temperature conditions were used to check for incompatibilities of the mAb with silicone oil, a necessary product contact material in PFS. In addition, screening studies to compare various closure materials as well as syringe barrel processing methods were used to select the optimum closure materials as well as the correct syringe processing method. Results indicate that the model mAb formulation used was sensitive to high levels of silicone oil especially under accelerated temperature conditions resulting in formation of protein–silicone particles in the solution for samples that were spiked with the silicone oil. Agitation stress did not have any significant impact on the quality attributes tested. Samples stored in syringe barrels that were processed with sprayed-on silicone had higher levels of subvisible particles as compared to those that were processed with the baked-on process. The tip cap comparability study resulted in one tip cap material having superior compatibility among the three that were tested. The quality attribute that was most impacted by the tip cap materials was mAb oxidation. An approach for evaluation of primary packaging components during the development of pre-filled syringe presentations for biotechnology-based compounds has been highlighted.