Purification and molecular characterization of a truncated-type Ara h 1, a major peanut allergen: oligomer structure, antigenicity, and glycoform

Asaduzzaman, Md.; Maeda, Megumi; Matsui, Teruaki; Takasato, Yoshihiro; Ito, Komei; Kimura, Yoshinobu

doi:10.1007/s10719-020-09969-1

Cited by 11 publications

(6 citation statements)

References 19 publications

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“…This study reported for the first time the successful selection of specific Nbs against peanut allergen of Ara h 1 and the development of Nb-pair based immunoassays. Ara h 1 is an allergen of 7S globulin belonging to the vicilin protein family with a molecular weight of around 65 kDa 24,32 and accounted for about 12% of peanut general protein. 33,34 For the preparation of Ara h 1, IEC was performed, and Ara h 1 protein with increased purity was obtained to ensure the successful screening of positive Nbs that specifically recognize Ara h 1 by comparing with application of unbiased strategies using crude protein extracts instead of the completely pure allergen.…”

Section: ■ Discussionmentioning

confidence: 99%

“…The initial purification of Ara h 1 was accomplished by using ion exchange chromatograph (IEC) and size exclusion chromatograph (SEC) based on its biochemical properties of isoelectric point (pI) and solubility. − In general, 20 g of peanut powder was resuspended in n -hexane (1:10, m/V ) to remove fat by stirring. The protein fraction was collected after centrifugation and concentrated to dry under nitrogen.…”

Section: Methodsmentioning

confidence: 99%

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Nanobody-Based Electrochemical Immunoassay for Sensitive Detection of Peanut Allergen Ara h 1

Lin

Peng

et al. 2023

J. Agric. Food Chem.

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Peanut is widely used for food supplementation with potential allergic reactions in infants and adults, which prompted the development of reliable and accurate detection of peanut allergens with emphasis on Ara h 1. In this study, a nanobody (Nb)-based micro-total electrochemical immunoassay (Nb-μTEI) was proposed to be generated. Generally, an alpaca was immunized with Ara h 1 to yield a Nb reservoir for selection of four specific Nbs. Nb-mediated immunocapturing allowed the identification of the target as Ara h 1. The Nb-based electrochemical immunoassay was developed by constructing a capturing electrode with cycles of signal enhancement. After construction of the capturing electrode, Nb152 with HA-tag was directly applied to connect immobilized anti-HA IgG for the capture of different concentrations of Ara h 1, which was labeled by biotinylated Nb152 to facilitate signal development with alkaline phosphatase conjugated streptavidin (SA-ALP). A linear range from 4.5 to 55 ng/mL was acquired with LOD and LOQ of 0.86 and 2.10 ng/mL, respectively, with an 11-fold increase of the sensitivity compared with the established sandwich ELISA. The dedicated immunoassay was verified by detecting the antigen spiked in food samples and demonstrated the successful conjugation of Nb with advanced detecting techniques.

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Section: ■ Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Nanobody-Based Electrochemical Immunoassay for Sensitive Detection of Peanut Allergen Ara h 1

Lin

Peng

et al. 2023

J. Agric. Food Chem.

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“…The main limitation of this type of expression system is the incorporation of glycans that are significantly different from human glycans, which hampers the immunological features of the produced VLPs (Lancaster et al, 2016). Extrinsic glycan signatures derived from nonhuman host platforms may provoke adverse effects like allergies or treatment rejection (Md et al, 2021). There are several factors that affect the production of HIV VLPs in insect cell lines with the baculovirus vector expression system, including multiplicity of infection (MOI), cell line, cell density and time of infection (Cervera et al, 2019).…”

Section: Major Developments On Hiv-1-based Vlp Productionmentioning

confidence: 99%

How promising are HIV-1-based virus-like particles for medical applications

Martins

Santos

Silva

et al. 2022

Front. Cell. Infect. Microbiol.

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New approaches aimed at identifying patient-specific drug targets and addressing unmet clinical needs in the framework of precision medicine are a strong motivation for researchers worldwide. As scientists learn more about proteins that drive known diseases, they are better able to design promising therapeutic approaches to target those proteins. The field of nanotechnology has been extensively explored in the past years, and nanoparticles (NPs) have emerged as promising systems for target-specific delivery of drugs. Virus-like particles (VLPs) arise as auspicious NPs due to their intrinsic properties. The lack of viral genetic material and the inability to replicate, together with tropism conservation and antigenicity characteristic of the native virus prompted extensive interest in their use as vaccines or as delivery systems for therapeutic and/or imaging agents. Owing to its simplicity and non-complex structure, one of the viruses currently under study for the construction of VLPs is the human immunodeficiency virus type 1 (HIV-1). Typically, HIV-1-based VLPs are used for antibody discovery, vaccines, diagnostic reagent development and protein-based assays. This review will be centered on the use of HIV-1-based VLPs and their potential biomedical applications.

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“…Although displaying specific exogenous glycan forms, like fungal glycans, can activate C‐type lectin receptors (CLRs) and promote an immune response (Lepenies et al, 2013 ), the use of biopharmaceuticals produced in nonhuman expression systems for clinical applications requires special attention. Exogenous immunologically active glycans may cause allergy reactions, therapy rejection, or other different side effects (Md et al, 2021 ; Tretter et al, 1993 ). Still within mammalian cell systems, nonhuman platforms like Chinese hamster ovary (CHO) cells present complex N ‐glycans capped with N ‐glycolylneuraminic acid (NeuGc), largely absent in human cells (Dhar et al, 2019 ; Ghaderi et al, 2010 ).…”

Section: Introductionmentioning

confidence: 99%

Differential N‐ and O‐glycosylation signatures of HIV‐1 Gag virus‐like particles and coproduced extracellular vesicles

Lavado‐García

Zhang

Cervera

et al. 2022

Biotech & Bioengineering

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Human immunodeficiency virus 1 (HIV-1) virus-like particles (VLPs) are nanostructures derived from the self-assembly and cell budding of Gag polyprotein.Mimicking the native structure of the virus and being noninfectious, they represent promising candidates for the development of new vaccines as they elicit a strong immune response. In addition to this, the bounding membrane can be functionalized with exogenous antigens to target different diseases. Protein glycosylation depends strictly on the production platform and expression system used and the displayed glycosylation patterns may influence downstream processing as well as the immune response. One of the main challenges for the development of Gag VLP production bioprocess is the separation of VLPs and coproduced extracellular vesicles (EVs). In this study, porous graphitized carbon separation method coupled with mass spectrometry was used to characterize the N-and O-glycosylation profiles of Gag VLPs produced in HEK293 cells. We identified differential glycan signatures between VLPs and EVs that could pave the way for further separation and purification strategies to optimize downstream processing and move forward in VLP-based vaccine production technology.

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Purification and molecular characterization of a truncated-type Ara h 1, a major peanut allergen: oligomer structure, antigenicity, and glycoform

Cited by 11 publications

References 19 publications

Nanobody-Based Electrochemical Immunoassay for Sensitive Detection of Peanut Allergen Ara h 1

Nanobody-Based Electrochemical Immunoassay for Sensitive Detection of Peanut Allergen Ara h 1

How promising are HIV-1-based virus-like particles for medical applications

Differential N‐ and O‐glycosylation signatures of HIV‐1 Gag virus‐like particles and coproduced extracellular vesicles

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