title: Toward Biomarker Development in Large Clinical Cohorts: An Integrated High-Throughput 96-Well-Plate-Based Sample Preparation Workflow for Versatile Downstream Proteomic Analyses.
abstract: We describe a cheap, robust, fast, high-throughput, and flexible proteomic sample processing method based on a regular 96-well plate by acetone precipitation under low centrifuge speed (96PACS), which enables predigestion processing of 96 samples within 2 h. Tested on a complex Huh-7 total lysate, 96PACS produced comparable proteome coverage and even showed better reproducibility than FASP. Quantitative performance of 96PACS was further tested using data-independent acquisition and parallel reaction monitoring quantitation in a set of 6 benchmark samples consisting of 6 serial dilutions of BSA spiked in complex E. coli proteome background. The protocol was also successfully modified for automation and was validated in a comparative label-free proteomic study to develop serum markers for early detection of liver fibrosis and necroinflammation in patients chronically infected with hepatitis B virus.
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University , 310003 Hangzhou, People's Republic of China.
authors.forename: Zeyu
authors.initials: Z
authors.lastname: Sun
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University , 310003 Hangzhou, People's Republic of China.
authors.forename: Xiaoli
authors.initials: X
authors.lastname: Liu
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University , 310003 Hangzhou, People's Republic of China.
authors.forename: Jing
authors.initials: J
authors.lastname: Jiang
authors.affiliation: Department of Infectious Diseases, Zhejiang Provincial People's Hospital , 310024 Hangzhou, Zhejiang, People's Republic of China.
authors.forename: Haijun
authors.initials: H
authors.lastname: Huang
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University , 310003 Hangzhou, People's Republic of China.
authors.forename: Jie
authors.initials: J
authors.lastname: Wang
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University , 310003 Hangzhou, People's Republic of China.
authors.forename: Daxian
authors.initials: D
authors.lastname: Wu
authors.affiliation: State Key Laboratory for Diagnosis and Treatment of Infectious Diseases, Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, The First Affiliated Hospital, College of Medicine, Zhejiang University , 310003 Hangzhou, People's Republic of China.
authors.forename: Lanjuan
authors.initials: L
authors.lastname: Li
mesh_terms: D015415:Biomarkers / Q000032:analysis; D015331:Cohort Studies; D029968:Escherichia coli Proteins / Q000032:analysis*; D006515:Hepatitis B virus / Q000302:isolation & purification; D057166:High-Throughput Screening Assays; D006801:Humans; D007249:Inflammation / Q000175:diagnosis / Q000821:virology; D008103:Liver Cirrhosis / Q000175:diagnosis / Q000821:virology; D040901:Proteomics
chemical_list: D015415:Biomarkers; D029968:Escherichia coli Proteins