Chapter title |
High-Throughput Parallel Proteomic Sample Preparation Using 96-Well Polyvinylidene Fluoride (PVDF) Membranes and C18 Purification Plates
|
---|---|
Chapter number | 27 |
Book title |
Serum/Plasma Proteomics
|
Published in |
Methods in molecular biology, July 2017
|
DOI | 10.1007/978-1-4939-7057-5_27 |
Pubmed ID | |
Book ISBNs |
978-1-4939-7056-8, 978-1-4939-7057-5
|
Authors |
Tue Bjerg Bennike, Hanno Steen |
Editors |
David W. Greening, Richard J. Simpson |
Abstract |
Meaningful proteomic-based biomarker discovery projects using primary human-derived specimens require the analysis of hundreds of samples in order to address the issue of interpersonal variability. Thus, robust high-throughput methods for the digestion of plasma samples are a prerequisite for such large clinical proteomic studies with hundreds of samples. Commonly used sample preparation methods are often difficult to parallelize and/or automate. Herein we describe a method for parallel 96-well plate-based sample preparation. Protein digestion is performed in 96-well polyvinylidene fluoride (PVDF) membrane plates and the subsequent purification in 96-well reversed phase C18 purification plates, enabling the usage of multichannel pipettes in all steps. The protocol can be applied using neat or depleted plasma/serum samples, but has also proven effective with other sample types. |
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