| Chapter title |
Phosphopeptide Enrichment and LC-MS/MS Analysis to Study the Phosphoproteome of Recombinant Chinese Hamster Ovary Cells
|
|---|---|
| Chapter number | 13 |
| Book title |
Heterologous Protein Production in CHO Cells
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6972-2_13 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6971-5, 978-1-4939-6972-2
|
| Authors |
Michael Henry, Orla Coleman, Prashant, Martin Clynes, Paula Meleady |
| Editors |
Paula Meleady |
| Abstract |
The reversible phosphorylation of proteins on serine, threonine, and tyrosine residues is one of the most important post-translational modifications that regulates many biological processes. The phosphoproteome has not been studied in any great detail in recombinant Chinese hamster ovary (CHO) cells to date despite phosphorylation playing a crucial role in regulating many molecular and cellular processes relevant to bioprocess phenotypes including, for example, transcription, translation, growth, apoptosis, and signal transduction. In this chapter, we provide a protocol for the phosphoproteomic analysis of Chinese hamster ovary cells using phosphopeptide enrichment with metal oxide affinity chromatography (MOAC) and immobilized metal affinity chromatography (IMAC) techniques, followed by site-specific identification of phosphorylated residues using LC-MS (MS2 and MS3) strategies. |
Mendeley readers
Geographical breakdown
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|---|---|---|
| Unknown | 15 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 3 | 20% |
| Student > Ph. D. Student | 2 | 13% |
| Unspecified | 1 | 7% |
| Student > Bachelor | 1 | 7% |
| Student > Master | 1 | 7% |
| Other | 1 | 7% |
| Unknown | 6 | 40% |
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|---|---|---|
| Biochemistry, Genetics and Molecular Biology | 5 | 33% |
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| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 7% |
| Unspecified | 1 | 7% |
| Unknown | 6 | 40% |