Proteohistography–Direct Analysis of Tissue with High Sensitivity and High Spatial Resolution Using ProteinChip Technology

On the proteomic level, all tissues, tissue constituents, or even single cells are heterogeneous, but the biological relevance of this cannot be adequately investigated with any currently available technique. The analysis of proteins of small tissue areas by any proteomic approach is limited by the number of required cells. Increasing the number of cells only serves to lower the spatial resolution of expressed proteins. To enhance sensitivity and spatial resolution we developed Proteohistography. Laser microdissection was used to mark special areas of interest on tissue sections attached to glass slides. These areas were positioned under microscopic control directly on an affinity chromatographic ProteinChip Array so that cells were lysed and their released proteins bound on a spatially defined point. The ProteinChip System, surface enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS), allows the laser to be steered to up to 215 distinct positions across the surface of the spot, enabling a high spatial resolution of measured protein profiles for the analyzed tissue area. Protein profiles of the single positions were visually plotted over the used tissue section to visualize distribution proteohistologically. Results show that the spatial distribution of detectable proteins could be used as a Proteohistogram for a given tissue area. Consequently, this procedure can provide additional information to both a matrix-assisted laser desorption/ionization (MALDI)-based approach and immunohistochemistry, as it is more sensitive, highly quantitative, and no specific antibody is needed. Hence, proteomic heterogeneity can be visualized even if proteins are not known or identified.

[1]  R. Bast,et al.  Three Biomarkers Identified from Serum Proteomic Analysis for the Detection of Early Stage Ovarian Cancer , 2004, Cancer Research.

[2]  D. Chan,et al.  Evaluation of serum protein profiling by surface-enhanced laser desorption/ionization time-of-flight mass spectrometry for the detection of prostate cancer: I. Assessment of platform reproducibility. , 2005, Clinical chemistry.

[3]  K. Lillemoe,et al.  Identification of hepatocarcinoma-intestine-pancreas/pancreatitis-associated protein I as a biomarker for pancreatic ductal adenocarcinoma by protein biochip technology. , 2002, Cancer research.

[4]  B. Rockhill.  Proteomic patterns in serum and identification of ovarian cancer , 2002, The Lancet.

[5]  E. Petricoin,et al.  Proteomic patterns in serum and identification of ovarian cancer , 2002, The Lancet.

[6]  Douglas C Pearl,et al.  Proteomic patterns in serum and identification of ovarian cancer , 2002, The Lancet.

[7]  U. Settmacher,et al.  Discovery and identification of alpha-defensins as low abundant, tumor-derived serum markers in colorectal cancer. , 2005, Gastroenterology.

[8]  Walter S. Liggett,et al.  Measurement Reproducibility in the Early Stages of Biomarker Development , 2005, Disease markers.

[9]  G. Wright,et al.  Development of a novel proteomic approach for the detection of transitional cell carcinoma of the bladder in urine. , 2001, The American journal of pathology.

[10]  E. Petricoin,et al.  Use of proteomic patterns in serum to identify ovarian Cancer , 2002 .

[11]  K. Kozak,et al.  Identification of biomarkers for ovarian cancer using strong anion-exchange ProteinChips: Potential use in diagnosis and prognosis , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[12]  L. Liotta,et al.  Proteomic Patterns of Nipple Aspirate Fluids Obtained by SELDI-TOF: Potential for New Biomarkers to Aid in the Diagnosis of Breast Cancer , 2002, Disease markers.

[13]  D. Driesch,et al.  Proteome Analysis of Maternal Serum Samples for Trisomy 21 Pregnancies Using ProteinChip Arrays and Bioinformatics , 2005, The journal of histochemistry and cytochemistry : official journal of the Histochemistry Society.

[14]  Mahlon D. Johnson,et al.  Protein Profiling in Brain Tumors Using Mass Spectrometry , 2004, Clinical Cancer Research.

[15]  T. Uchida,et al.  Application of a novel protein biochip technology for detection and identification of rheumatoid arthritis biomarkers in synovial fluid. , 2002, Journal of proteome research.

[16]  Georgia Lahr,et al.  Identification of expressed genes by laser-mediated manipulation of single cells , 1998, Nature Biotechnology.

[17]  Yu Shyr,et al.  Proteomic patterns of tumour subsets in non-small-cell lung cancer , 2003, The Lancet.

[18]  S. Gygi,et al.  Haptoglobin-alpha subunit as potential serum biomarker in ovarian cancer: identification and characterization using proteomic profiling and mass spectrometry. , 2003, Clinical cancer research : an official journal of the American Association for Cancer Research.

[19]  F. von Eggeling,et al.  Biomarker Discovery and Identification in Laser Microdissected Head and Neck Squamous Cell Carcinoma with ProteinChip® Technology, Two-dimensional Gel Electrophoresis, Tandem Mass Spectrometry, and Immunohistochemistry* , 2003, Molecular & Cellular Proteomics.

[20]  Uwe Claussen,et al.  Mass spectrometry meets chip technology: A new proteomic tool in cancer research? , 2001, Electrophoresis.

[21]  S. Gygi,et al.  Haptoglobin-α subunit as potential serum biomarker in ovarian cancer: Identification and characterization using proteomic profiling and mass spectrometry , 2003 .

[22]  S. Weinberger,et al.  Protein quantification by the SELDI-TOF-MS–based ProteinChip® System , 2005 .

[23]  Roy A Jensen,et al.  Proteomics in diagnostic pathology: profiling and imaging proteins directly in tissue sections. , 2004, The American journal of pathology.

[24]  H. Davies,et al.  Tissue-specific microdissection coupled with ProteinChip array technologies: applications in cancer research. , 2000, BioTechniques.

[25]  Y. Kato,et al.  Rapid discovery and identification of a tissue-specific tumor biomarker from 39 human cancer cell lines using the SELDI ProteinChip platform. , 2003, Biochemical and biophysical research communications.

[26]  Günther Ernst,et al.  A Technical Triade for Proteomic Identification and Characterization of Cancer Biomarkers , 2004, Cancer Research.

[27]  T. Yip,et al.  New desorption strategies for the mass spectrometric analysis of macromolecules , 1993 .