Optimisation of Total Protein Concentration Based on Buffer Type and Ratio in Placental Protein Extraction of Bovine Kedah Kelantan

• Main Author: Tee Yung Yee
• Format: Undergraduate Final Project Report
• Language: English
• Published: 2019
• Online Access: http://discol.umk.edu.my/id/eprint/4848/1/TEE%20YUNG%20YEE.pdf; http://discol.umk.edu.my/id/eprint/4848/

Proteomic study related with placental protein has been an interest topic to the researcher over decades to determine the function and mechanism of particular protein that perhaps to improve pregnancy related disorder or pregnancy diagnosis in human and livestock industry. Protein extraction is a preliminary step of protein purification which mainly focus on maximisation of total protein yield. The heterogenous properties causes diversification of protein, therefore there is no absolute protocol in protein extraction, the type of buffer and ratio could give different protein concentration in different type of mammalian tissue, hence lead to the study of optimisation of types of buffer and buffer ratio to obtain better total protein yield. The objectives of this study are to compare the total protein yield based on three types of buffer with three different ratios. Three types of buffer included phosphate buffer saline (PBS), radioimmunoprecipitation assay (RIPA) buffer and RIPA buffer with addition of protease inhibitor (Pi) whilst sample to buffer ratio, 1:1, 1:3, and 1:5 were used in the series of protein extraction process that involved mechanical disruption, incubation, sonication, and centrifμgation. Bradford assay was carried out to determine the total protein yield based on the standard curve of bovine serum albumin (BSA). From this study it is concluded there is a significant interaction between buffer type and ratio (P<0.05) where the use RIPA buffer with 1:1 ratio gave the best total protein yield (194.880 ± 15.089 mg/g). This study is perhaps to be a useful reference to future study in term of protein extraction of mammalian tissue especially placental tissue.