開發針對炎症標誌物單體C反應蛋白之禽類單鏈抗體

Development of Avian Single-Chain Variable Fragment Antibody Targeting the Inflammatory Biomarker Monomeric C-Reactive Protein

單體C反應蛋白（mCRP）在血小板凝集、嗜中性球趨化以及促炎性細胞因子釋放等發炎相關反應中扮演關鍵角色，突顯其作為診斷與治療標的之潛力與重要性。本研究中，我們利用純化之mCRP蛋白免疫雞隻，並自雞蛋中純化獲得多株IgY抗體，經由Western blot與ELISA驗證其對mCRP具專一性免疫反應。進一步地，我們以噬菌體展示技術建立短鏈與長鏈兩種scFv抗體基因庫，經四輪生物篩選後，從表現良好的抗體中隨機挑選26株進行定序，結果可分為五類抗體。其中三株scFv抗體（cCLPS2、cCLPL1與cCLPL2）在Western blot與ELISA分析中皆顯示具mCRP專一性結合能力。這些單鏈抗體為發炎相關疾病之診斷與治療提供了新的可能，未來將進一步評估其於臨床應用之潛力。

Monomeric C-reactive protein (mCRP) plays a critical role in inflammation related responses such as platelet aggregation, neutrophil chemotaxis, and the release of pro-inflammatory cytokines, highlighting its potential and importance as a diagnostic and therapeutic target. In this study, chickens were immunized with purified mCRP protein, and polyclonal IgY antibodies were subsequently purified from egg yolks. The specificity of the IgY antibodies against mCRP was confirmed by Western blot and ELISA. Furthermore, two scFv antibody libraries, including short linker and long linker, were constructed using phage display technology. After four rounds of bio-panning, 26 clones were randomly selected from expressed clones for sequencing, revealing five distinct antibody groups. Among them, three scFv antibodies (cCLPS2, cCLPL1, and cCLPL2) demonstrated specific binding to mCRP on both Western blot and ELISA analyses. These antibodies offer new opportunities for the diagnosis and treatment of inflammation related diseases, and their potential clinical applications will be further evaluated in future studies.