Technological aspects of manufacturing and analytical control of biological nanoparticles

Brezgin, Sergey; Parodi, Alessandro; Kostyusheva, Anastasiya; Ponomareva, N. I.; Lukashev, Alexander N.; Sokolova, Darina V.; Pokrovsky, Vadim S.; Slatinskaya, O. V.; Максимов, Г. В.; Zamyatnin, Andrey A.; Chulanov, Vladimir; Kostyushev, Dmitry

doi:10.1016/j.biotechadv.2023.108122

Cited by 26 publications

(17 citation statements)

References 225 publications

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“…[34] TFF uses a semi-permeable membrane and a pressure gradient to effectively remove unwanted contaminants while retaining desired EVs. [35] The TFF process offers significant advantages for handling large volumes of biological samples, as evidenced by prior research in diverse fields including bioprocessing and biopharmaceutical manufacturing. [34, 36] In conjunction with TFF, SEC further enhances the isolation process by enabling the separation of EVs based on their hydrodynamic radius.…”

Section: Discussionmentioning

confidence: 99%

Scalable Isolation of Surface-Engineered Extracellular Vesicles and Separation of Free Proteins via Tangential Flow Filtration and Size Exclusion Chromatography (TFF-SEC)

Kawai-Harada,

Nimmagadda,

Harada

2024

Preprint

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Extracellular vesicles (EVs) represent small lipid bilayer structures pivotal in mediating intercellular communication via biomolecular transfer.1Their inherent characteristics, including packaging, non-immunogenicity, and biofluid stability, position EVs as promising drug delivery vectors. However, developing clinical quality EVs requires multifaceted technological advancement. In this study, a method is introduced for engineering extracellular vesicles (eEVs) from cultured cells and their subsequent isolation using lab-scale tangential flow filtration (TFF). Initially, cells are transfected with EV-display constructs to facilitate the secretion of eEVs bearing the desired coding molecules. Following brief centrifugation, the cell culture media undergoes filtration using hollow fiber filters. TFF, by applying a constant flow, effectively segregates molecules based on designated molecular weight cut-off (MWCO), enriching particles between 50 nm and 650 nm. Compared to conventional methods like ultracentrifugation, TFF demonstrates higher efficiency in removing undesired molecules/aggregates while exerting less stress on EVs. Characterization of eEVs through various assays confirms TFF’s superiority in isolating pure EV populations. Additionally, the necessity of size-exclusion chromatography (SEC) after tangential flow filtration (TFF) becomes evident for effectively removing unbound protein contaminants. In conclusion, TFF-SEC emerges as a scalable and superior approach for eEV isolation, promising significant advancements in clinical applications.

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Section: Discussionmentioning

confidence: 99%

Scalable Isolation of Surface-Engineered Extracellular Vesicles and Separation of Free Proteins via Tangential Flow Filtration and Size Exclusion Chromatography (TFF-SEC)

Kawai-Harada,

Nimmagadda,

Harada

2024

Preprint

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show abstract

“…UF, compared to UC, offers lower costs and faster processing and does not require specialized equipment [27]. However, loss of EVs may occur due to membrane attachment, vesicle entrapment, or filter media clogging [28]. Nanomembrane concentrators with brief centrifugation periods are also effective alternatives to UF for isolating urinary exosomes from cell-free samples such as urine, serum, cerebrospinal fluid, and cell culture medium [27].…”

Section: Size-based Methodsmentioning

confidence: 99%

Analysis and Characterization of the Extracellular Vesicles Released in Non-Cancer Diseases Using Matrix-Assisted Laser Desorption Ionization/Mass Spectrometry

Aresta,

De Vietro,

Zambonin

2024

IJMS

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The extracellular vesicles (EVs) released by cells play a crucial role in intercellular communications and interactions. The direct shedding of EVs from the plasma membrane represents a fundamental pathway for the transfer of properties and information between cells. These vesicles are classified based on their origin, biogenesis, size, content, surface markers, and functional features, encompassing a variety of bioactive molecules that reflect the physiological state and cell type of origin. Such molecules include lipids, nucleic acids, and proteins. Research efforts aimed at comprehending EVs, including the development of strategies for their isolation, purification, and characterization, have led to the discovery of new biomarkers. These biomarkers are proving invaluable for diagnosing diseases, monitoring disease progression, understanding treatment responses, especially in oncology, and addressing metabolic, neurological, infectious disorders, as well as advancing vaccine development. Matrix-Assisted Laser Desorption Ionization (MALDI)/Mass Spectrometry (MS) stands out as a leading tool for the analysis and characterization of EVs and their cargo. This technique offers inherent advantages such as a high throughput, minimal sample consumption, rapid and cost-effective analysis, and user-friendly operation. This review is mainly focused on the primary applications of MALDI–time-of-flight (TOF)/MS in the analysis and characterization of extracellular vesicles associated with non-cancerous diseases and pathogens that infect humans, animals, and plants.

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“…Fourthly, the specificity of exosomes needs to be further enhanced. Surface modification is an effective way to improve the specificity of exosome delivery by using exosomal surface proteins as affinity tags and modifying specific proteins onto the exosomal surface [ 181 ]. Surface modification has been successfully used to enhance exosome targeting to brain cells and improve their ability to cross the blood–brain barrier [ 182 ].…”

Section: Potential Applications Of Exosomes In Hepatitis Bmentioning

confidence: 99%

Exosomes target HBV-host interactions to remodel the hepatic immune microenvironment

Wu,

Niu,

Shi

2024

J Nanobiotechnol

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Chronic hepatitis B poses a significant global burden, modulating immune cells, leading to chronic inflammation and long-term damage. Due to its hepatotropism, the hepatitis B virus (HBV) cannot infect other cells. The mechanisms underlying the intercellular communication among different liver cells in HBV-infected individuals and the immune microenvironment imbalance remain elusive. Exosomes, as important intercellular communication and cargo transportation tools between HBV-infected hepatocytes and immune cells, have been shown to assist in HBV cargo transportation and regulate the immune microenvironment. However, the role of exosomes in hepatitis B has only gradually received attention in recent years. Minimal literature has systematically elaborated on the role of exosomes in reshaping the immune microenvironment of the liver. This review unfolds sequentially based on the biological processes of exosomes: exosomes’ biogenesis, release, transport, uptake by recipient cells, and their impact on recipient cells. We delineate how HBV influences the biogenesis of exosomes, utilizing exosomal covert transmission, and reshapes the hepatic immune microenvironment. And based on the characteristics and functions of exosomes, potential applications of exosomes in hepatitis B are summarized and predicted. Graphical Abstract

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Technological aspects of manufacturing and analytical control of biological nanoparticles

Cited by 26 publications

References 225 publications

Scalable Isolation of Surface-Engineered Extracellular Vesicles and Separation of Free Proteins via Tangential Flow Filtration and Size Exclusion Chromatography (TFF-SEC)

Scalable Isolation of Surface-Engineered Extracellular Vesicles and Separation of Free Proteins via Tangential Flow Filtration and Size Exclusion Chromatography (TFF-SEC)

Analysis and Characterization of the Extracellular Vesicles Released in Non-Cancer Diseases Using Matrix-Assisted Laser Desorption Ionization/Mass Spectrometry

Exosomes target HBV-host interactions to remodel the hepatic immune microenvironment

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