Production, purification and characterization of chicken IgY directed to human hemoglobin

• Main Authors: Feng-So Chan, 詹豐碩
• Other Authors: Yu-Chie Wang
• Format: Others
• Language: zh-TW
• Published: 2014
• Online Access: http://ndltd.ncl.edu.tw/handle/74332551412145565804

碩士 === 國立臺灣師範大學 === 生命科學研究所 === 102 === Colorectal cancer (CRC) is one of the most prevalent cancers in the world. Routine fecal occult blood tests (FOBT) help detect CRC in its early stages and thus decrease CRC mortality and incidence. The aim of this study is to establish a production platform, using chickens as experimental animals, for the core component of anti-human hemoglobin antibody (anti-hHb Ab) used in the immunochemical FOBT (I-FOBT). To that end, we have examined the effects of antigen dose, adjuvant and booster injection on chicken’s humoral immune response and monitored changes of antibody titers in the sera and yolksfrom that the optimal parameters for anti-hHb Ab production can be deduced. As for antibody purification, we have prepared affinity column with antigen-coupled beads and developed an effective cleaning procedure for that column. The affinity-purified sample of anti-hHb IgY does not cross-react with non-hemoglobin proteins and exhibits superior purity and reactivity to antigen than that of commercially available. In addition, preliminary assessments have been performed to evaluate the sensitivity of using anti-hHb IgY in a biochemical assay and to scrutinize the plausible interferences exerted by animal blood-containing food or by upper gastrointestinal tract bleeding. In conclusion, the use of chickens for anti-hHb Ab production brings many advantages over the conventional mammal hosts, including lower cost, higher yield, easier handling process and greater compliance to animal welfare regulations. Since the genarated anti-hHb IgY reveals great sensitivity and specificity in biochemical assays, they can be considered as an excellent alternative for the rabbit anti-hHb IgG currently used in the commercial I-FOBT kits.