Nanoparticle-protein complexes mimicking corona formation in ocular environment

Jo, D.H.,Kim, J.H.,Son, J.G.,Dan, K.S.,Song, S.H.,Lee, T.G.,Kim, J.H. IPC Science and Technology Press 2016 Biomaterials Vol.109 No.-

Nanoparticles adsorb biomolecules to form corona upon entering the biological environment. In this study, tissue-specific corona formation is provided as a way of controlling protein interaction with nanoparticles in vivo. In the vitreous, the composition of the corona was determined by the electrostatic and hydrophobic properties of the associated proteins, regardless of the material (gold and silica) or size (20- and 100-nm diameter) of the nanoparticles. To control protein adsorption, we pre-incubate 20-nm gold nanoparticles with 5 selectively enriched proteins from the corona, formed in the vitreous, to produce nanoparticle-protein complexes. Compared to bare nanoparticles, nanoparticle-protein complexes demonstrate improved binding to vascular endothelial growth factor (VEGF) in the vitreous. Furthermore, nanoparticle-protein complexes retain in vitro anti-angiogenic properties of bare nanoparticles. In particular, priming the nanoparticles (gold and silica) with tissue-specific corona proteins allows nanoparticle-protein complexes to exert better in vivo therapeutic effects by higher binding to VEGF than bare nanoparticles. These results suggest that controlled corona formation that mimics in vivo processes may be useful in the therapeutic use of nanomaterials in local environment.

Enzyme delivery using protein-stabilizing and cell-penetrating 30Kc19α protein nanoparticles

Park, Hee Ho,Woo, Yeon Hwa,Ryu, Jina,Lee, Hong Jai,Park, Ju Hyun,Park, Tai Hyun Elsevier Science B.V., Amsterdam. 2017 PROCESS BIOCHEMISTRY Vol.63 No.-

<P><B>Abstract</B></P> <P>Nanoparticles (NPs) are an emerging strategy for drug delivery and have been studied for the delivery of various biomolecules, such as chemically synthesized drugs and therapeutic proteins. In particular, protein NPs are non-cytotoxic and biodegradable. Application of a full length recombinant 30Kc19 protein to human serum albumin (HSA) NPs has been shown to improve the cellular uptake and stability of the cargo enzyme. In this study, we demonstrate that drug delivery can be achieved with only the α-helix domain of the 30Kc19 protein (30Kc19α), and without the addition of HSA. Protein concentration and pH were crucial for NP generation. NPs had a uniformly spherical shape with an optimal diameter of 180–230nm, and released β-galactosidase in a sustained manner. The 30Kc19α protein provided stability to the cargo enzyme, and helped maintain the specific activity of the enzyme. X-gal staining showed effective delivery of β-galactosidase into human dermal fibroblasts. Non-cytotoxic property of the 30Kc19α protein demonstrates that such NPs could be a resourceful tool for delivering drugs to cells.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Protein nanoparticles can be generated with the α-helix domain of 30Kc19. </LI> <LI> Human serum albumin is not required for 30Kc19α protein nanoparticle generation. </LI> <LI> 30Kc19α nanoparticles showed high intracellular delivery potential for large cargos. </LI> <LI> 30Kc19α protein provided stability and helped maintain specific activity of enzyme. </LI> </UL> </P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Protein corona: a new approach for nanomedicine design

Nguyen, Van Hong,Lee, Beom-Jin Dove Medical Press 2017 INTERNATIONAL JOURNAL OF NANOMEDICINE Vol.12 No.-

<P>After administration of nanoparticle (NP) into biological fluids, an NP–protein complex is formed, which represents the “true identity” of NP in our body. Hence, protein–NP interaction should be carefully investigated to predict and control the fate of NPs or drug-loaded NPs, including systemic circulation, biodistribution, and bioavailability. In this review, we mainly focus on the formation of protein corona and its potential applications in pharmaceutical sciences such as prediction modeling based on NP-adsorbed proteins, usage of active proteins for modifying NP to achieve toxicity reduction, circulation time enhancement, and targeting effect. Validated correlative models for NP biological responses mainly based on protein corona fingerprints of NPs are more highly accurate than the models solely set up from NP properties. Based on these models, effectiveness as well as the toxicity of NPs can be predicted without in vivo tests, while novel cell receptors could be identified from prominent proteins which play important key roles in the models. The ungoverned protein adsorption onto NPs may have generally negative effects such as rapid clearance from the bloodstream, hindrance of targeting capacity, and induction of toxicity. In contrast, controlling protein adsorption by modifying NPs with diverse functional proteins or tailoring appropriate NPs which favor selective endogenous peptides and proteins will bring promising therapeutic benefits in drug delivery and targeted cancer treatment.</P>

Targeted cellular delivery of robust enzyme nanoparticles for the treatment of drug-induced hepatotoxicity and liver injury

Lee, Min Sang,Kim, Nak Won,Lee, Jung Eun,Kim, Myung Goo,Yin, Yue,Kim, Sun Young,Ko, Bo Sung,Kim, Aeseon,Lee, Jong Han,Lim, Su Yeon,Lim, Dong Woo,Kim, Sun Hwa,Park, Ji Won,Lim, Yong Taik,Jeong, Ji Hoon Elsevier 2018 Acta Biomaterialia: structure-property-function re Vol.81 No.-

<P><B>Abstract</B></P> <P>Direct delivery of proteins into cells has been considered an effective approach for treating the protein-related diseases. However, clinical use of proteins has still been limited due to their instability in the blood and poor membrane permeability. To achieve an efficient cellular delivery of the protein to target cells via a systemic administration, a multifunctional carrier system having desirable stability both in the blood stream and the cells, specific cell-targeting property and endosomal escape functions may be required. In this study, we prepared a catalytic nanoparticle containing an active enzyme by cross-tethering multiple superoxide dismutase (SOD) molecules with catechol-derivatized hyaluronic acid (HA). The permeable shell of hydrophilic HA chains effectively protects the enzyme from degradation in the blood after intravenous administration and provides an additional function for targeting hepatocytes expressing HA receptor (CD44). The structure and catalytic activity of the enzyme molecules in the nanoparticle were not significantly compromised in the nanoparticle. In addition, ultra-small calcium phosphate nanoparticles (USCaP, 2–5 nm) were crystalized and decorated on the surface of the nanoparticle for the efficient endosomal escape after cellular uptake. The SOD-containing nanoparticle fortified with USCaP was used for the treatment of acetaminophen (APAP)-induced fulminant hepatotoxicity and liver injury. The nanoparticle achieved the efficient hepatic cellular delivery of SOD via a systemic administration and resulted in efficient removal of reactive oxygen species (ROS) in the liver and remarkable improvement of APAP-induced hepatotoxicity and liver injury in animals.</P> <P><B>Statement of Significance</B></P> <P>Despite the enormous therapeutic potential, the intracellular delivery of proteins has been limited due to their poor membrane permeability and stability. In this study, we demonstrated an active enzyme-containing nanoparticle functionalized by hyaluronic acid and ultra-small size calcium phosphate nanoparticles (2–5 nm) for targeted cellular delivery of superoxide dismutase (SOD). The nanoparticle was designed to integrate all the essential functions, including serum stability, target specificity, and endosomal escape capability, for a systemic delivery of a therapeutic protein to the cells of the liver tissue. The intravenous administration of the nanoparticle efficiently removes reactive oxygen species (ROS) in the liver and remarkably improves the drug-induced hepatotoxicity and the progress of fulminant liver injury in an acetaminophen-overdose animal model.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

단백질 나노입자를 이용한 약물전달 시스템

홍세영(Seyoung Hong),박희호(Hee Ho Park) 한국생물공학회 2020 KSBB Journal Vol.35 No.1

Nanoparticles have various advantages as drug carriers, so they have been widely used to deliver various chemical and biomolecule drugs, such as anti-cancer drugs and therapeutic proteins. Among nanoparticles, protein nanoparticles are non-cytotoxic and biodegradable, with high potential as drug carriers. Protein nanoparticles can be synthesized using proteins such as silk protein fibroin, albumin, gelatin, gliadin, legumin, 30Kc19 protein derived from silkworm hemolymph, and are manufactured using methods such as emulsion/solvent extension, desolvation, electrospray technology, complexification, and etc. The characterization parameters of protein nanoparticles are particle size, particle morphology and structure, surface charge, drug loading and release, which vary the drug delivery efficiency according to these parameters. Thus, the synthesis of protein nanoparticles using various materials and manufacturing techniques is being actively studied to improve drug delivery efficiency.

Protein nanoparticles directed cancer imaging and therapy

Miao Yao,Yang Tao,Yang Shuxu,Yang Mingying,Mao Chuanbin 나노기술연구협의회 2022 Nano Convergence Vol.9 No.2

Cancer has been a serious threat to human health. Among drug delivery carriers, protein nanoparticles are unique because of their mild and environmentally friendly preparation methods. They also inherit desired characteristics from natural proteins, such as biocompatibility and biodegradability. Therefore, they have solved some problems inherent to inorganic nanocarriers such as poor biocompatibility. Also, the surface groups and cavity of protein nanoparticles allow for easy surface modification and drug loading. Besides, protein nanoparticles can be combined with inorganic nanoparticles or contrast agents to form multifunctional theranostic platforms. This review introduces representative protein nanoparticles applicable in cancer theranostics, including virus-like particles, albumin nanoparticles, silk protein nanoparticles, and ferritin nanoparticles. It also describes the common methods for preparing them. It then critically analyzes the use of a variety of protein nanoparticles in improved cancer imaging and therapy.

콜로이드 골드 나노입자의 단백질 수송성 평가법

김미영,노상명,김정목,최한곤,김정애,오유경 한국약제학회 2004 Journal of Pharmaceutical Investigation Vol.34 No.6

Colloidal gold nanoparticles might be of use as nano scale delivery systems of various therapeutic materials in the firture. Recent studies have reported the feasibility of colloidal gold nanoparticles as gene delivery systems or protein delivery systems. In this study, we aimed to develop a short-step method useful for screening the optimal coating conditions of colloidal gold nanoparticles with proteins. We observed that colloidal gold nanoparticles have properties of changing its unique color when they were exposed to NaCl solution. Taking advantage of the color changing properties of colloidal gold nano-particles, we applied the color testing method of colloidal gold nanoparticles solutions for evaluating the protein coating nature. Using bovine senim albumin as a model protein, we tested the protein coating of colloidal gold nanoparticles via the color change upon NaCl addition. The optimal coating concentration and coating conditions of colloidal gold nanoparticles with bovine serum albumin were fixed using the color testing methods. We suggest that the color testing method might be applied to optimize the coating condition of colloidal gold nanoparticles with other therapeutic proteins.

Cloaking nanoparticles with protein corona for targeted drug delivery

김채규,유자형 한국공업화학회 2018 한국공업화학회 연구논문 초록집 Vol.2018 No.1

Translating nanoparticle-based agents into clinical applications still remains a challenge due to the difficulty in regulating interactions occurring at the interfaces between nanoparticles and biological systems. Upon contact with physiological environments, a nanoparticle interacts biomolecules, induces a protein layer on its surface, and complicates its biological effects, resulting in the loss of the targeting specificity and rapid clearance via phagocyte system. In this talk, we present a new targeting strategy for nanoparticles incorporated with a pre-coated protein corona shield. The protein corona shield featuring recombinant fusion proteins generically combined HER2- binding affibody molecules prevented non-specific interactions with serum proteins while ensuring systematic targeting functions in vitro and in vivo. This study provides a new insight into the use of the protein corona as a targeting agent with specificity.

His-tagged protein immobilization on cationic ferrite magnetic nanoparticles

박성진,김승연,김승훈,박경민,황병희 한국화학공학회 2018 Korean Journal of Chemical Engineering Vol.35 No.6

Magnetic nanoparticles have been applied in various fields because of their interesting magnetic properties. Immobilization on magnetic nanoparticles is a very important step in functionalizing them. We examined protein immobilization efficiency using interactions between his-tagged enhanced green fluorescence protein and affordable cationic ferrite magnetic nanoparticles for the first time. Four types of ferrite magnetic nanoparticles were verified: cobalt iron oxide, copper iron oxide, nickel iron oxide, and iron (III) oxide as negative controls. Among the four ferrite magnetic nanoparticles, copper ferrite magnetic nanoparticle was confirmed to have the highest immobilization efficiency at 3.0mg proteins per gram ferrite magnetic nanoparticle and 78% of total enhanced green fluorescence protein. In addition, the maximum binding efficiency was determined for copper ferrite magnetic nanoparticle. Consequently, this newly verified his-tag-immobilizing capacity of copper ferrite magnetic nanoparticle could provide a facile, capable, and promising strategy for immobilizing his-tagged proteins or peptides with high purity for biosensors, magnetic separation, or diagnostics.

Protein corona on magnetite nanoparticles and internalization of nanoparticle-protein complexes into healthy and cancer cells

Jiang, Wen,Lai, Kuilin,Wu, Yao,Gu, Zhongwei 대한약학회 2014 Archives of Pharmacal Research Vol.37 No.1

Superparamagnetic magnetite nanoparticles (MNPs) of different surface properties are incubated in complicated living fluid, including fetal bovine serum solution, cell complete culture medium and cell culture system with/without serum, to investigate the alteration of protein corona and its impact on cell internalization. The MNPs prepared by co-precipitation method are functionalized with L-Lysine (Lys), Glucosamic acid (GA) to obtain amine, carboxyl and hydroxyl groups, separately. All the particles adsorb serum proteins to form MNPs-protein complexes with the surface charge changing into negative. 1D SDS/PAGE gel images analysis indicates that the composition and content of hard protein corona on the surface of NPs are related to their functional groups and agglomeration, and the total amount of protein in the medium. In cell culture system, particles not only adsorb serum proteins, but also associate with cytosolic proteins arising from HepG2 and L02 cells. GA modified MNPs (MNPs-GA) exhibit bovine serum albumin anti-adsorption capability because of the terminal hydroxyl and carboxyl groups. MNPs-GA also shows the highest cellular uptake and label efficiency compared with uncoated MNPs and Lys modified MNPs, due to larger aggregates formation and specific protein corona composition, rather than commonly approved electrostatic interaction between particles and cells. For the first time, our results provide visualized reports on previously neglected, but indispensable protein corona of the MNPs after interaction with both healthy and cancer cells, suggesting that cytosolic protein corona from cells and aggregation of particles are important factors needed to be account for on studying the nano-bio interface.