Publication: Optimization and Standardization of Thermal Treatment as a Plasma Prefractionation Method for Proteomic Analysis
5
Issued Date
2019-01-01
Resource Type
ISSN
23146141
23146133
23146133
Other identifier(s)
2-s2.0-85066070161
Rights
Mahidol University
Rights Holder(s)
SCOPUS
Bibliographic Citation
BioMed Research International. Vol.2019, (2019)
Suggested Citation
Wararat Chiangjong, Channarong Changtong, Jirawan Panachan, Churat Weeraphan, Chantragan Srisomsap, Suradej Hongeng, Jisnuson Svasti, Somchai Chutipongtanate Optimization and Standardization of Thermal Treatment as a Plasma Prefractionation Method for Proteomic Analysis. BioMed Research International. Vol.2019, (2019). doi:10.1155/2019/8646039 Retrieved from: https://repository.li.mahidol.ac.th/handle/123456789/50392
Research Projects
Organizational Units
Authors
Journal Issue
Thesis
Title
Optimization and Standardization of Thermal Treatment as a Plasma Prefractionation Method for Proteomic Analysis
Abstract
© 2019 Wararat Chiangjong et al. Prefractionation is a prerequisite step for deep plasma proteomics. Highly abundant proteins, particularly human serum albumin (HSA) and immunoglobulin G (IgG), typically interfere with investigation of proteins with lower abundance. A relatively simple preparation method based on high temperature can precipitate thermolabile proteins, providing a strategic window to access the thermostable plasma subproteome. This study aimed to optimize thermal treatment as a reliable prefractionation method and to compare it with two commercial kits, including HSA and IgG immunodepletion (IMDP) and combinatorial peptide ligand libraries (CPLL), using untreated plasma as a control condition. By varying the temperature and the incubation period, the optimal condition was found as treatment at 95°C for 20 min, which maintained about 1% recovery yield of soluble proteins. Consistency and reproducibility of thermal treatment-derived plasma subproteome were checked by two-dimensional electrophoresis. The coefficient of variation regarding protein spot numbers was less than 10% among three independent specimens. Highly abundant protein depletion of the thermal treatment was evaluated by immunoblotting against HSA and IgG as compared to the untreated plasma, IMDP, and CPLL. Multidimensional comparison based on 489 unique peptides derived from the label-free quantitative mass spectrometry revealed that the thermal treatment, IMDP, and CPLL provided distinct sets of plasma subproteome compared to untreated plasma, and these appeared to be complementary to each other. Comparing the characteristics of the three procedures suggested that thermal treatment was more cost-effective and less time-consuming than IMDP and CPLL. This study proposes the use of thermal treatment as a reliable and cost-effective method for plasma prefractionation which provides benefits to large-scale proteomic projects and biomarker studies.