To produce recombinant Arachis hypogaea Allergen Ara h 1, clone P17, the first step involves isolating the gene encoding the 26-614aa of the Allergen Ara h 1, clone P17. This gene is cloned into an appropriate expression vector along with an N-terminal 6xHis-tag gene. The vector is introduced into an E.coli cell via transformation. Inside the host cell, the target gene is expressed, leading to the production of the recombinant protein. The protein is harvested from the cell lysate and purified often using affinity chromatography. The final step is to determine the purity of this recombinant Arachis hypogaea Allergen Ara h 1, clone P17 by SDS-PAGE. Its purity is greater than 90%.
Arachis hypogaea Allergen Ara h 1, clone P17 is a variant of the major peanut allergen Ara h 1 found in peanuts, which along with Ara h 2 and Ara h 3, contributes significantly to peanut allergies [1][2][3]. The allergenicity of Ara h 1 has been linked to the specific arrangement of monomers in the homotrimeric structure of vicilin/7S globulin proteins [4]. Peanut allergies, mediated by immunoglobulin E (IgE) sensitization to various proteins including Ara h allergens, are a significant and potentially life-threatening health concern.
References:
[1] K. Hilu, S. Friend, V. Vallanadu, L. Hollingsworth, & D. Bevan, Molecular evolution of genes encoding allergen proteins in the peanuts genus arachis: structural and functional implications, Plos One, vol. 14, no. 11, p. e0222440, 2019. https://doi.org/10.1371/journal.pone.0222440
[2] A. Barre, C. Nguyen, C. Granier, H. Benoist, & P. Rougé, Ige-binding epitopes of pis v 1, pis v 2 and pis v 3, the pistachio (pistacia vera) seed allergens, Allergies, vol. 1, no. 1, p. 63-91, 2021. https://doi.org/10.3390/allergies1010006
[3] M. Shen, A. Joshi, R. Vannam, C. Dixit, R. Hamilton, C. Kumaret al., Epitope‐resolved detection of peanut‐specific ige antibodies by surface plasmon resonance imaging, Chembiochem, vol. 19, no. 3, p. 199-202, 2018. https://doi.org/10.1002/cbic.201700513
[4] L. Lange, K. Beyer, & J. Kleine-Tebbe, Benefits and limitations of molecular diagnostics in peanut allergy, Allergo Journal International, vol. 23, no. 5, p. 158-163, 2014. https://doi.org/10.1007/s40629-014-0019-z
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