Premium
Proteomic approach to human kidney glomerulus prepared by laser microdissection from frozen biopsy specimens: Exploration of proteome after removal of blood‐derived proteins
Author(s) -
Yoshida Yutaka,
Nameta Masaaki,
Kuwano Masayoshi,
Zhang Ying,
Bo Xu,
Magdeldin Sameh,
Cui Zenyui,
Fujinaka Hidehiko,
Yaoita Eishin,
Tomonaga Takeshi,
Yamamoto Tadashi
Publication year - 2012
Publication title -
proteomics – clinical applications
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.948
H-Index - 54
eISSN - 1862-8354
pISSN - 1862-8346
DOI - 10.1002/prca.201200016
Subject(s) - laser capture microdissection , proteome , microdissection , proteomics , biopsy , frozen section procedure , blood proteins , pathology , glomerulus , renal glomerulus , kidney , chemistry , biology , microbiology and biotechnology , biochemistry , medicine , glomerulonephritis , gene expression , gene , endocrinology
Purpose Abundance of blood‐derived proteins in glomeruli prepared by laser microdissection from human kidney biopsy specimens has hampered in‐depth proteomic analysis of glomeruli. We attempted to establish experimental platform for in‐depth proteomic analysis of glomeruli by removal of blood‐derived proteins from frozen biopsy samples. Experimental design Frozen sections of biopsy samples were exposed to repeated PBS washes prior to laser microdissection to remove blood‐derived proteins, and glomerular dissectants were analyzed by MS. The depth of proteomic analysis was evaluated by dynamic range of identified proteins and detection of low‐abundance proteins. Results Two times PBS washes of frozen sections effectively eliminated blood‐derived proteins in laser‐microdissected glomeruli and gave an increased number of identified proteins. Analysis of glomeruli from single specimens by a linear ion trap‐Orbitrap mass analyzer generated nonredundant, high‐confidence datasets of more than 400 identified proteins with high reproducibility, which attained to a considerable depth of the glomerulus proteome as revealed by a wide dynamic range and identification of low‐abundance proteins. Conclusions and clinical relevance Implementation of washing of frozen section with PBS successfully removed blood‐derived proteins and resulted in an in‐depth proteomic analysis of laser‐microdissected glomeruli, suggesting applicability to clinical study.