Isolation, characterization, and immunoinformatics of ige-mediated Group III (Trypsin-like Serine Protease) allergen from house dust mite suidasia pontifica Bornea, Richard Christopher C., Calica, Patricia Nicole H., Dela Cruz, John Wayne R., Isidro, Khayie Nicole M., Malit, Erika Pauline B. 6

Undergraduate Thesis: (BS Biology major in Cell and Molecular Biology) - Pamantasan ng Lungsod ng Maynila, 2024 56

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ABSTRACT: Allergy is an understudied and underestimated medical condition that affects roughly 30-50% of the human population through allergy derived symptoms as well as psychological discomfort due to deterioration of the affected individual's quality of life. Development of long-term medical solutions through the knowledge of its presence, origin, and prognosis are hindered by unmet needs in medical training, education, and basic allergology research. Yet its devastating effect can reach its victims even without outdoor exposure via indoor sources, which excessively worsen asthma and other symptoms chronically without the patient's awareness of need for medical intervention. One of the major sources of these indoor allergens emerge though fecal constituents of dust mite species under the Order Sarcoptiformes. Though assays can be performed through skin prick tests, it is unfortunate that the characterization of allergens from the molecular level still lacks data that can provide a means of acquiring pure allergens for downstream medical pipelines and assessment, especially in the Philippines. Therefore, this study aims to isolate and identify tropical dust mite species Suidasia pontifica and molecularly characterize one of its expected, yet undiscovered group 3 allergen. The methodological approach includes the transcriptomic extraction of S. pontifica Trypsin-like serine protease mRNA and subsequent amplification through RT-PCR. The extracted mRNA was documented for gel electrophoresis and the bands were extracted for single-pass sanger sequencing. Sequence data was annotated, modeled, and compared to other medically important Sarcoptiformes. Results present the successful isolation and morphological identification of S. pontifica species from San Pedro Laguna. Sequenced data produced an mRNA similar to group 3 allergen of other species. Annotation and Immunoinformatics discovered homology on B-cell, T-cell and Allergenic motifs across the entire Sui p 3 sequence. It is also suggested to perform in vitro study of the allergen for actual determination of immune response and allergenic potential.

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