Production and Functional Analysis of Monoclonal Antibodies Against Proline-rich Peptide

• Main Authors: Huang, Shih-Han, 黃詩涵
• Other Authors: Wu, Fu-Yu
• Format: Others
• Language: zh-TW
• Published: 2013
• Online Access: http://ndltd.ncl.edu.tw/handle/e986c8

碩士 === 國立宜蘭大學 === 動物科技學系碩士班 === 101 === Alzheimer's disease (AD) is a neurodegenerative disease, the most common dementia in aged people. As lifespan increases globally, AD attract more and more attentions. The pathogenic factor for AD is β-amyloid (Aβ), which accumulated in brain and formed fibrous plagues. Proline-rich peptides (PRP) is originally extracted from sheep colostrum, and contain amino acid sequence homology with β-casein. Studies have shown that the PRP improve cognitive function in AD patients, and can inhibit Aβ aggregation. PRP is already in the market with the trademark of ColostrininTM. The requirement of colostrum as source material limits the quantity of production, and jeopardizes the health of the newborn animals. This study was to generate monoclonal antibody against PRP for isolation of PRP and further research. PRP were synthesized based on the amino acid sequence of β-casein, and were conjugated with KLH for mice immunization. Alternatively, PRP were synthesized as multiple antigen peptide systems (MAPS). After increase of serum antibody titer as measured by ELISA, hybridoma was performed. A few cell lines with positive reaction to PRP have been screened out. Far-dot blot was used to detect if PRP can react with Aβ. Briefly, Aβ was doted on NC membrane, blocked with BSA, incubated with MAPS-PRP, and then detected with monoclonal antibody against PRP. The result showed that PRP can react with Aβ. For the future research, PRP monoclonal antibody will be used to make affinity chromatography for rapid purification of PRP, and to study the functions of PRP.