TY - JOUR
T1 - MALDI imaging mass spectrometry of N-linked glycans on formalin-fixed paraffin-embedded murine kidney
AU - Gustafsson, Ove J R
AU - Briggs, Matthew T.
AU - Condina, Mark R.
AU - Winderbaum, Lyron J.
AU - Pelzing, Matthias
AU - McColl, Shaun R.
AU - Everest-Dass, Arun V.
AU - Packer, Nicolle H.
AU - Hoffmann, Peter
N1 - Copyright the Author(s) 2014. Version archived for private and non-commercial use with the permission of the author/s and according to publisher conditions. For further rights please contact the publisher.
PY - 2015/3
Y1 - 2015/3
N2 - Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies.
AB - Recent developments in spatial proteomics have paved the way for retrospective in situ mass spectrometry (MS) analyses of formalin-fixed paraffin-embedded clinical tissue samples. This type of analysis is commonly referred to as matrix-assisted laser desorption/ionization (MALDI) imaging. Recently, formalin-fixed paraffin-embedded MALDI imaging analyses were augmented to allow in situ analyses of tissue-specific N-glycosylation profiles. In the present study, we outline an improved automated sample preparation method for N-glycan MALDI imaging, which uses in situ PNGase F-mediated release and measurement of N-linked glycans from sections of formalin-fixed murine kidney. The sum of the presented data indicated that N-glycans can be cleaved from proteins within formalin-fixed tissue and characterized using three strategies: (i) extraction and composition analysis through on-target MALDI MS and liquid chromatography coupled to electrospray ionization ion trap MS; (ii) MALDI profiling, where N-glycans are released and measured from large droplet arrays in situ; and (iii) MALDI imaging, which maps the tissue specificity of N-glycans at a higher resolution. Thus, we present a complete, straightforward method that combines MALDI imaging and characterization of tissue-specific N-glycans and complements existing strategies.
UR - http://www.scopus.com/inward/record.url?scp=84964694274&partnerID=8YFLogxK
UR - http://purl.org/au-research/grants/arc/LP110100693
U2 - 10.1007/s00216-014-8293-7
DO - 10.1007/s00216-014-8293-7
M3 - Article
C2 - 25434632
AN - SCOPUS:84964694274
SN - 1618-2642
VL - 407
SP - 2127
EP - 2139
JO - Analytical and bioanalytical chemistry
JF - Analytical and bioanalytical chemistry
IS - 8
ER -