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Expression and Characterization of Functional Recombinant Bet v 1.0101 in the Chloroplast of Chlamydomonas reinhardtii.
Hirschl, Sonja; Ralser, Claudia; Asam, Claudia; Gangitano, Alessandro; Huber, Sara; Ebner, Christof; Bohle, Barbara; Wolf, Martin; Briza, Peter; Ferreira, Fatima; Griesbeck, Christoph; Wallner, Michael.
Affiliation
  • Hirschl S; Department of Biotechnology and Food Engineering, MCI Management Center Innsbruck, Innsbruck, Austria.
Int Arch Allergy Immunol ; 173(1): 44-50, 2017.
Article in En | MEDLINE | ID: mdl-28494467
BACKGROUND: Allergen immunotherapy (AIT) still plays a minor role in the treatment of allergic diseases. To improve the acceptance of AIT by allergic patients, the treatment has to become more convenient and efficacious. One possibility is the oral application of allergens or derivatives thereof. Therefore, we sought to produce a recombinant allergen in the green alga Chlamydomonas reinhardtii as a novel production platform. METHODS: The major birch pollen allergen Bet v 1 was selected as candidate molecule, and a codon-optimized gene was synthesized and stably integrated into the microalga C. reinhardtii FUD50. Positive transformants were identified by PCR, cultured, and thereafter cells were disrupted by sonication. Bet v 1 was purified from algal total soluble protein (TSP) by affinity chromatography and characterized physicochemically as well as immunologically. RESULTS: All transformants showed expression of the allergen with yields between 0.01 and 0.04% of TSP. Algal-derived Bet v 1 displayed similar secondary structure elements as the Escherichia coli-produced reference allergen. Moreover, Bet v 1 produced in C. reinhardtii showed binding comparable to human IgE as well as murine Bet v 1-specific IgG. CONCLUSION: We could successfully produce recombinant Bet v 1 in C. reinhardtii. As microalgae are classified as GRAS (generally recognized as safe), the pilot study supports the development of novel allergy treatment concepts such as the oral administration of allergen-containing algal extracts for therapy.
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Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Plant Proteins / Recombinant Proteins / Allergens / Chloroplasts / Chlamydomonas reinhardtii / Antigens, Plant Limits: Humans Language: En Journal: Int Arch Allergy Immunol Journal subject: ALERGIA E IMUNOLOGIA Year: 2017 Document type: Article Affiliation country: Country of publication:

Full text: 1 Collection: 01-internacional Database: MEDLINE Main subject: Plant Proteins / Recombinant Proteins / Allergens / Chloroplasts / Chlamydomonas reinhardtii / Antigens, Plant Limits: Humans Language: En Journal: Int Arch Allergy Immunol Journal subject: ALERGIA E IMUNOLOGIA Year: 2017 Document type: Article Affiliation country: Country of publication: