E-ISSN 2146-8362
 

Original Research 


The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue

Pumza Magangane, Nonhlanhla Khumalo, Henry Adeola.

Abstract
Abstract
Objectives: The success of novel in-situ immunohistoproteomics approaches that employ Matrix-assisted laser desorption and ionization (MALDI) mass spectrometry imaging (MSI) heavily depends on good antigen retrieval efficacy.
Methods: We have optimized the effect of various antigen retrieval buffers on peptide profiles acquired from FFPE traction alopecia scalp tissue. Heat-induced epitope retrieval (HIER) was performed using Citrate, Tris, and Tris-EDTA buffers under various conditions. MALDI-MSI analysis of the tissue was performed using a high-throughput, state-of-the-art RapiFlex MALDI TOF/TOF tissue-typer equipped with a smart beam laser.
Results: Background matrix peaks were found under most conditions except under Tris-EDTA buffer. Trypsin digestion periods longer than overnight and possibly higher concentrations are needed for Citrate and Tris. The Tris-EDTA buffer performed best for MALDI-MSI antigen retrieval at an incubation period of overnight at 60C.
Conclusions: We have identified in this study that antigen retrieval conditions need to be optimised prior to MALDI MSI analysis of different tissue types.

Key words: Skin, MALDI MSI, antigen retrieval, buffer, mass spectrometry, FFPE


 
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How to Cite this Article
Pubmed Style

Magangane P, Khumalo N, Adeola H. The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue. J Interdiscip Histopathol. 2018; 6(2): 26-32. doi:10.5455/jihp.20180528101354


Web Style

Magangane P, Khumalo N, Adeola H. The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue. http://www.ejmjih.com/?mno=298794 [Access: July 17, 2018]. doi:10.5455/jihp.20180528101354


AMA (American Medical Association) Style

Magangane P, Khumalo N, Adeola H. The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue. J Interdiscip Histopathol. 2018; 6(2): 26-32. doi:10.5455/jihp.20180528101354



Vancouver/ICMJE Style

Magangane P, Khumalo N, Adeola H. The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue. J Interdiscip Histopathol. (2018), [cited July 17, 2018]; 6(2): 26-32. doi:10.5455/jihp.20180528101354



Harvard Style

Magangane, P., Khumalo, N. & Adeola, H. (2018) The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue. J Interdiscip Histopathol, 6 (2), 26-32. doi:10.5455/jihp.20180528101354



Turabian Style

Magangane, Pumza, Nonhlanhla Khumalo, and Henry Adeola. 2018. The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue. Journal of Interdisciplinary Histopathology, 6 (2), 26-32. doi:10.5455/jihp.20180528101354



Chicago Style

Magangane, Pumza, Nonhlanhla Khumalo, and Henry Adeola. "The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue." Journal of Interdisciplinary Histopathology 6 (2018), 26-32. doi:10.5455/jihp.20180528101354



MLA (The Modern Language Association) Style

Magangane, Pumza, Nonhlanhla Khumalo, and Henry Adeola. "The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue." Journal of Interdisciplinary Histopathology 6.2 (2018), 26-32. Print. doi:10.5455/jihp.20180528101354



APA (American Psychological Association) Style

Magangane, P., Khumalo, N. & Adeola, H. (2018) The effect of antigen retrieval buffers on MALDI mass spectrometry imaging of peptide profiles in skin FFPE tissue. Journal of Interdisciplinary Histopathology, 6 (2), 26-32. doi:10.5455/jihp.20180528101354





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