TY - JOUR T1 - A novel ELISA-based approach for quantitative analysis of biomolecules in formalin-fixed paraffin-embedded tissues TT - Formalinle fikslenmiş parafin gömülü dokularda biyomoleküllerin kantitatif analizi için yeni formül AU - Türeli, Serkan AU - Kuloğlu, Onur Alp AU - Karakoç, Elif AU - Karahan, Sevilay AU - Gököz, Özay AU - Usubütün, Alp AU - Atilla, Pergin PY - 2025 DA - October Y2 - 2025 JF - Journal of Medicine and Palliative Care JO - J Med Palliat Care / JOMPAC / Jompac PB - MediHealth Academy Yayıncılık WT - DergiPark SN - 2717-7505 SP - 447 EP - 451 VL - 6 IS - 5 LA - en AB - Aims: Enzyme-linked immunosorbent assay (ELISA) is a widely used method for protein quantification in biological specimens. However, its application to formalin-fixed paraffin-embedded (FFPE) tissue is limited due to inconsistent results and a lack of standardization. We aimed to develop and apply a novel formula to standardize ELISA results derived from FFPE tissue samples and to assess its feasibility using inflammatory skin disease biopsies.Methods: FFPE skin tissue blocks from individuals diagnosed with psoriasis (n=6), cutaneous lupus erythematosus (CLE, n=6), lichen sclerosis (LS, n=6), and healthy controls (n=6) were included. Tissue homogenates were prepared and tumor necrosis factor-α (TNF- α) levels were measured using a commercial ELISA kit. A new calculation formula was developed, normalizing absorbance values to protein concentration and biopsy size. Results: Application of the new formula enabled more consistent quantification of the TNF-α levels across heterogenous sized FFPE specimens. TNF- α levels were significantly elevated in all patients group compared to the controls (p KW - ELISA KW - FFPE tissue KW - TNF-alpha KW - skin biopsy KW - inflammation KW - cytokine N2 - Amaçlar: Enzim bağlantılı immünosorbent test (ELISA), biyolojik örneklerde protein miktarının ölçülmesinde yaygın olarak kullanılan bir yöntemdir. Ancak, formalinle fikse edilmiş parafin bloklanmış (FFPE) dokulara uygulanması, tutarsız sonuçlar ve standardizasyon eksikliği nedeniyle sınırlıdır. Bu çalışmada, FFPE doku örneklerinden elde edilen ELISA sonuçlarını standardize etmek amacıyla yeni bir formül geliştirmeyi ve inflamatuar deri hastalığı biyopsileri üzerinde uygulanabilirliğini değerlendirmeyi amaçladık.Yöntemler: Psoriasis (n = 6), kutanöz lupus eritematozus (CLE, n = 6), liken sklerozus (LS, n = 6) tanısı konmuş bireyler ile sağlıklı kontrollerden (n = 6) elde edilen FFPE deri doku blokları çalışmaya dahil edildi. Doku homojenatları hazırlandı ve tümör nekroz faktörü-α (TNF-α) düzeyleri ticari bir ELISA kiti kullanılarak ölçüldü. Absorbans değerlerini protein konsantrasyonu ve biyopsi boyutuna göre normalize eden yeni bir hesaplama formülü geliştirildi.Bulgular: Yeni formülün uygulanması, farklı boyutlardaki FFPE örneklerinde TNF-α düzeylerinin daha tutarlı şekilde ölçülmesini sağladı. TNF-α düzeyleri tüm hasta gruplarında kontrol grubuna kıyasla anlamlı derecede yüksekti (p < 0.05). Ayrıca, CLE örneklerindeki TNF-α konsantrasyonu, psoriasis ve LS örneklerinden anlamlı olarak daha yüksekti.Sonuçlar: Bu çalışma, FFPE örneklerinde tekrarlanabilir ELISA kantifikasyonu sağlayan yeni ve standardize bir formül sunmaktadır. Söz konusu formül, retrospektif ve arşiv doku temelli araştırmalarda pratik bir araç olarak kullanılabilir. CR - Tabatabaei MS, Ahmed M. Enzyme-linked immunosorbent assay (ELISA). Methods Mol Biol. 2022;2508:115-134. doi:10.1007/978-1-0716-2376-3_10 CR - Lin AV. Direct ELISA. 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