@article{article_1758433, title={A novel ELISA-based approach for quantitative analysis of biomolecules in formalin-fixed paraffin-embedded tissues}, journal={Journal of Medicine and Palliative Care}, volume={6}, pages={447–451}, year={2025}, author={Türeli, Serkan and Kuloğlu, Onur Alp and Karakoç, Elif and Karahan, Sevilay and Gököz, Özay and Usubütün, Alp and Atilla, Pergin}, keywords={ELISA, FFPE doku, TNF-alfa, Deri biyopsisi, İnflamasyon, Sitokin}, abstract={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 <0.05), TNF-α concentration in CLE samples was significantly higher than in psoriasis and LS samples. Conclusion: This study introduces a novel, standardized formula that allows reproducible ELISA quantification in FFPE specimens. It may serve as a practical tool in retrospective and archival tissue-based research.}, number={5}, publisher={MediHealth Academy Yayıncılık}, organization={Hacettepe University Scientific Research Projects Coordination Unit (THD-2022-19666)}