Analytical Proteomics Research Group (CI8)
Main lines of research:
- Use of high-resolution capillary electrophoresis with UV detection diode beam as an analytical technique for carrying out the study of the plasma proteome.
- Development and implementation of strategies for the selection of profiles of biomarkers that help in early detection of metastatic prostate cancer.
For this, public microarray databases were used and bioinformatics tools for the selection of a profile of genes associated with metastatic disease. Subsequently, proteomic analysis was performed on cell lines of prostate cancer, an androgen-dependent (LNCaP) and other androgen-resistant (PC3), to identify biomarkers encoded by genes previously selected. It was also developed sample pretreatment strategies based on immunoprecipitation, using a new pattern of magnetic particles to carry out selective and specific extraction of target proteins, using antibodies. The above immunoprecipitation was applied for the pre-treatment of cell samples using these antibodies. After protein identification by Western blot techniques etc, we proceeded to the development and optimization of liquid chromatography techniques coupled to mass spectrometry for the analysis of target proteins. We developed this project with the collaboration of the Proteomics Group of Mass Spectrometry Unit, University of California, San Francisco (UCSF) led by Prof. AL Burlingame.
- A third research project in Clinical Proteomics, funded by the National Institutes of Health (NIH), USA,
It focuses on the study of the phosphoproteome of lymphocytes, the project is being developed in collaboration with the University of California, San Francisco (UCSF ), USA, pioneer group in Proteomics with Mass Spectrometry. The development and optimization of enrichment techniques based mainly on various modes of affinity chromatography and development, optimization and application of isotopic labeling techniques such as iTRAQ for quantification of proteins present.
Immunochemistry and in particular immunoaffinity, has been a development tool widely used in the research lines developed by this group, especially in regard to their use as sample preparation strategies for the extraction and purification of analytes present in complex matrices. The European project "Immunoprobes for Food Contamination Analysis," provided funds, in which Dr. Ana Gago Martinez was Coordinator for the Spanish side.
The draft tools were developed for solid phase extraction, based on immunoaffinity which formed the basis for future strategies based on the same principle. The limitations encountered due in most cases, to the unspecificity of the antibodies used, led researchers to consider an investigation based on the development of "chemosynthetic" antibodies that will be useful to achieve adequate specificity, which will allow us to have a specific tool for isolation (extraction, purification, etc.) of the analyte of interest. It will also constitute a high potential tool for proteomic analysis, especially for the extraction and enrichment of minority proteins present in complex matrices such as cell tissue.
The Project will provide antibodies chemically modified with various application purposes (therapeutic, analytical, etc. ..) and it is being done with the collaboration of Dr. W. Robert Arathoon former Vice President of Genentech, California, renowned expert in the field of antibody technology).
The group led by Prof. Ana Gago Martínez has broad experience in the development and optimization of analytical methodologies for the treatment of complex biological samples. Combining separation techniques (chromatography and electrophoresis) with detection ones, Mass Spectrometry, for the determination and confirmation of analytes present in matrices of high complexity. Special emphasis has been placed on the development of sample preparation strategies to improved selectivity and sensitivity, necessary in the treatment of these matrices and subsequent analysis of various compounds present at the level of traces or subtraces.
The application of these methodologies to aquatic biotoxins has resulted in a dense scientific production in high impact journals, as well as invitations to conferences of international relevance in the area of marine biotoxins, and funding in various European, national and regional projects. international recognition to this achievements was reflected in naming Dr Ana Gago Martinez as Spanish Representative to vote in the "TASK FORCE ON MARINE AND FRESHWATER TOXINS" of the AOAC, with focus on the evaluation of analytical methods developed worldwide and subsequently validated prior to its adoption as official methods of analysis by the AOAC.
All work done in developing analytical methods to control aquatic biotoxins, and the subsequent evaluation of their toxicity, have provided the basis to extend the methodologies developed into other areas. In this case the experience in dealing with complex matrices and the search for analytes with similar characteristics was instrumental in initiating research work in proteomics.