2009
DOI: 10.1007/978-1-59745-493-3_20
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Analysis of Protein Glycosylation and Phosphorylation Using Liquid Phase Separation, Protein Microarray Technology, and Mass Spectrometry

Abstract: Summary Protein glycosylation and phosphorylation are very common posttranslational modifications. The alteration of these modifications in cancer cells is closely related to the onset and progression of cancer and other disease states. In this protocol, strategies for monitoring the changes in protein glycosylation and phosphorylation in serum or tissue cells on a global scale and specifically characterizing these alterations are included. The technique is based on lectin affinity enrichment for glycoproteins… Show more

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Cited by 12 publications
(9 citation statements)
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“…Current techniques used in biopharmaceutical processing are typically suited to only monitoring a few sample characteristics simultaneously, require lengthy sample preparation, or are costly. For example, mass spectrometry (MS), is usually focussed at the molecular level to monitor post translational modifications (PTMs), and for the identification of numerous proteins in a protein mixture, but it is costly to run [40]. UV spectroscopy is utilised in a variety of set ups to elucidate protein-protein interactions through monitoring changes in UV absorbance using delta absorbance (ΔAbs), but is used primarily for protein concentration measurements [41][42][43].…”
Section: Ftir Spectroscopy and Spectroscopic Imagingmentioning
confidence: 99%
“…Current techniques used in biopharmaceutical processing are typically suited to only monitoring a few sample characteristics simultaneously, require lengthy sample preparation, or are costly. For example, mass spectrometry (MS), is usually focussed at the molecular level to monitor post translational modifications (PTMs), and for the identification of numerous proteins in a protein mixture, but it is costly to run [40]. UV spectroscopy is utilised in a variety of set ups to elucidate protein-protein interactions through monitoring changes in UV absorbance using delta absorbance (ΔAbs), but is used primarily for protein concentration measurements [41][42][43].…”
Section: Ftir Spectroscopy and Spectroscopic Imagingmentioning
confidence: 99%
“…Typically, M6P levels can be assessed by either mass spectrometry [15] or by an anion exchange chromatography method that requires a pulsed amperometric detector [16]. Either of these approaches involves processing of the protein samples by multiples steps before quantification.…”
Section: Resultsmentioning
confidence: 99%
“…26 The advantage of a glycoprotein-lectin array is that the glycoproteins do not require labeling, while the disadvantage is that the high effective concentration of the samples within one spot increases the tendency for spurious interactions. 27,28 In the present discussion, we emphasize the fabrication and detection techniques of the normal phase lectin microarrays (Fig. 1(a)-(d)).…”
Section: Lectin Microarray Platformmentioning
confidence: 99%