Abstract
Allergic disease diagnosis is currently experiencing a breakthrough due to the use of allergenic molecules in serum-based assays rather than allergen extracts in skin tests. The former methodology is considered a very innovative technology compared with the latter, since it is characterized by flexibility and adaptability to the patient’s clinical history and to microtechnology, allowing multiplex analysis. Molecular-based analysis requires pure allergens to detect IgE sensitization, and a major goal, to maintain the diagnosis cost-effective, is to limit their production costs. In addition, for the production of recombinant eukaryotic proteins similar to natural ones, plant-based protein production is preferred to bacterial-based systems due to its ability to perform most of the post-translational modifications of eukaryotic molecules. In this framework, Plant Molecular Farming (PMF) may be useful, being a production platform able to produce complex recombinant proteins in short time-frames at low cost. As a proof of concept, PMF has been exploited for the production of Bet v 1a, a major allergen associated with birch (Betula verrucosa) pollen allergy. Bet v 1a has been produced using two different transient expression systems in Nicotiana benthamiana plants, purified and used in a new generation multiplex allergy diagnosis system, the patient-Friendly Allergen nano-BEad Array (FABER). Plant-made Bet v 1a is immunoreactive, binding IgE and inhibiting IgE-binding to the Escherichia coli expressed allergen currently available in the FABER test, thus suggesting an overall similar though non-overlapping immune activity compared with the E. coli expressed form.
Highlights
Allergic diseases, defined as abnormal responses of the human body when in contact with an allergen, have become a common health problem worldwide, with a rising incidence both in adults and children (Lau et al, 2018)
For the infection with the Potato Virus X (PVX)-based vector, the N. benthamiana leaves were infiltrated with the A. tumefaciens suspension carrying pGPVXGATEWAY(A).Betv1
The development of tools for a precise and definitive diagnosis of allergic diseases is considered strategic for the future, both in terms of developing vaccines for immunotherapy, exploiting a companion diagnostic strategy, and of studying the genuine allergic sesitization of patients to a particular allergen source
Summary
Allergic diseases, defined as abnormal responses of the human body when in contact with an allergen, have become a common health problem worldwide, with a rising incidence both in adults and children (Lau et al, 2018). It is rather difficult to obtain exact epidemiology data on this topic but it has been estimated that 25% of the general population suffer from these diseases (Sampson, 2004). The primary diagnostic tool currently used for allergic disease diagnosis is skin prick testing (SPT), which evaluates the presence and degree of cutaneous reactivity against a surrogate marker of sensitization, composed of protein extracts from allergenic sources. This method is hampered by several problems.
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