Expression of grape class IV chitinase in Spodoptera frugiperda (Sf9) insect cells

Falak, Reza; Varasteh, Abdolreza; Ketabdar, Hanieh; Sankian, Mojtaba

doi:10.1016/j.aller.2012.11.004

Cited by 5 publications

(5 citation statements)

References 38 publications

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“…Meanwhile, comparable IgE reactivity of the recombinant and native forms suggest that some post-translational modifications, such as glycosylation, do not contribute to parvalbumin allergenicity. Similar findings were also reported for other allergens (Falak et al, 2014;Rosmilah, Shahnaz, Masita, Noormalin, & Jamaludin, 2005). Therefore, expression in prokaryotic systems, which usually do not provide glycosylation, did not abolish IgE binding of wolf-herring parvalbumin.…”

Section: Discussionsupporting

confidence: 84%

“…Proteins were resolved on an AKTA Prime Plus FPLC system (GE healthcare) using a diethylaminoethyl (DEAE) sepharose CL-6B column as the anion exchange matrix and eluted at a flow rate of 1 ml/min by a linear gradient mixture of starting buffer (20 mM Tris-HCl pH 8.6) supplemented with 1 M NaCl. Following SDS-PAGE, the desired fractions were selected for further studies (Falak et al, 2013;Falak, Varasteh, Ketabdar, & Sankian, 2014).…”

Section: Parvalbumin Purificationmentioning

confidence: 99%

“…Insoluble material was removed by centrifugation at 14,000 × g for 30 min. Recombinant parvalbumin was purified from the soluble phase of the lysate (supernatant) by metal affinity chromatography (Ni-IDA agarose, ParsToos, Iran) according to the manufacturer's instructions (Falak et al, 2014).…”

Section: Cloning and Expression Of Parvalbumin In Prokaryotic Systemmentioning

confidence: 99%

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Expression of recombinant parvalbumin from wolf-herring fish and determination of its IgE-binding capability

Mohammadi

Mokhtarian

Kardar

et al. 2017

Food and Agricultural Immunology

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In this study, we produced the recombinant form of parvalbumin from wolf-herring fish and determined its IgE reactivity. Parvalbumin cDNA was sub-cloned into pET28 and expressed in Escherichia coli BL-21. The immunoreactivities of the recombinant and native parvalbumins were compared, and the effect of calcium binding was determined by sera from 25 fish-allergic patients. ELISA and Western blotting confirmed similar IgEreactivities of the recombinant and native proteins and confirmed that this phenomenon is highly dependent on calcium binding. The recombinant protein was 94.5% similar to carp parvalbumin (Cyp c1). Approximately 72% of patients reacted strongly with recombinant parvalbumin, 80% of them reacted with the native form and only 56% showed IgE reactivity with crude extract. Because the IgE-binding capacity of recombinant wolf-herring parvalbumin is retained and is highly similar to Cyp c1, the wild and hypoallergenic forms of this allergen could be used for diagnosis and immunotherapy of fish allergy, respectively. ARTICLE HISTORY

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Section: Discussionsupporting

confidence: 84%

Section: Parvalbumin Purificationmentioning

confidence: 99%

Section: Cloning and Expression Of Parvalbumin In Prokaryotic Systemmentioning

confidence: 99%

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Expression of recombinant parvalbumin from wolf-herring fish and determination of its IgE-binding capability

Mohammadi

Mokhtarian

Kardar

et al. 2017

Food and Agricultural Immunology

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“…Finally, recombinant protein expression was triggered by addition of 1 mM isopropyl-thiogalactoside (IPTG, Sigma, USA) and incubation of the bacterial suspension for 4–6 hours at 37°C. Subsequently, the bacterial cells were harvested by centrifugation and following cell lysis, rSs FAR was purified using a Hi-Trap chelating Sepharose column (Ni-NTA agarose, Qiagen, Hilden, Germany) [47].…”

Section: Methodsmentioning

confidence: 99%

Fatty acid and retinol-binding protein: A novel antigen for immunodiagnosis of human strongyloidiasis

et al. 2019

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The tenacious human parasitic helminth Strongyloides stercoralis is a significant health problem worldwide. The current lack of a definitive diagnostic laboratory test to rule out this infection necessitates designing more specific diagnostic methods. Fatty acid and retinol-binding protein (FAR) plays a crucial role in the development and reproduction of nematodes. We generated a recombinant form of this protein and determined its applicability for immunodiagnosis of S . stercoralis . The L3 form of S . stercoralis was harvested and used for RNA extraction and cDNA synthesis. The coding sequence of S . stercoralis FAR (SsFAR) was cloned into pET28a(+) vector, expressed in E . coli BL21 and purified. ELISA and immunoblotting were employed to determine the specificity and sensitivity of rSsFAR using a set of defined sera. In addition, we analyzed the phylogenetic relationship of SsFAR with different FAR sequences from other nematodes. The cloned SsFAR had an open reading frame of 447 bp encoding 147 amino acids, with a deduced molecular mass of 19 kD. The SsFAR amino acid sequence was 93% identical to FAR of S . ratti . For differential immunodiagnosis of strongyloidiasis, rSsFAR exhibited 100% sensitivity and 97% specificity. However, cross-reactivity with FAR proteins of other parasites, namely Toxocara canis and Echinococcus granulosus , was noted. Our results provide a novel approach for immunodiagnosis of S . stercoralis infections using rSsFAR with reliable sensitivity and specificity.

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“…One grape chitinase, including its chitin binding module, was expressed in insect cells [35]. It resulted allergenic, as demonstrated by immunoblot assay against the sera of patients allergic to grape.…”

Section: Class IVmentioning

confidence: 99%

Chitinases as Food Allergens

et al. 2019

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Food allergies originate from adverse immune reactions to some food components. Ingestion of food allergens can cause effects of varying severity, from mild itching to severe anaphylaxis reactions. Currently there are no clues to predict the allergenic potency of a molecule, nor are cures for food allergies available. Cutting-edge research on allergens is aimed at increasing information on their diffusion and understanding structure-allergenicity relationships. In this context, purified recombinant allergens are valuable tools for advances in the diagnostic and immunotherapeutic fields. Chitinases are a group of allergens often found in plant fruits, but also identified in edible insects. They are classified into different families and classes for which structural analyses and identification of epitopes have been only partially carried out. Moreover, also their presence in common allergen databases is not complete. In this review we provide a summary of the identified food allergenic chitinases, their main structural characteristics, and a clear division in the different classes.

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Expression of grape class IV chitinase in Spodoptera frugiperda (Sf9) insect cells

Cited by 5 publications

References 38 publications

Expression of recombinant parvalbumin from wolf-herring fish and determination of its IgE-binding capability

Expression of recombinant parvalbumin from wolf-herring fish and determination of its IgE-binding capability

Fatty acid and retinol-binding protein: A novel antigen for immunodiagnosis of human strongyloidiasis

Chitinases as Food Allergens

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