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Tuyển tập báo cáo các nghiên cứu khoa học quốc tế ngành hóa học dành cho các bạn yêu hóa học tham khảo đề tài: Immunoreactivity of anti-gelsolin antibodies: implications for biomarker validation | Haverland et al. Journal of Translational Medicine 2010 8 137 http www.translational-medicine.eom content 8 1 137 TRANSLATIONAL MEDICINE RESEARCH Open Access Immunoreactivity of anti-gelsolin antibodies implications for biomarker validation Nicole Haverland Gwenael Pottiez Jayme Wiederin Pawel Ciborowski Abstract Background Proteomic-based discovery of biomarkers for disease has recently come under scrutiny for a variety of issues one prominent issue is the lack of orthogonal validation for biomarkers following discovery. Validation by ELISA or Western blot requires the use of antibodies which for many potential biomarkers are under-characterized and may lead to misleading or inconclusive results. Gelsolin is one such biomarker candidate in HIV-associated neurocognitive disorders. Methods Samples from human plasma and CSF monkey plasma monocyte-derived macrophage supernatants and commercial gelsolin recombinant and purified were quantitated using Western blot assay and a variety of anti-gelsolin antibodies. Plasma and CSF was used for immunoaffinity purification of gelsolin which was identified in eight bands by tandem mass spectrometry. Results Immunoreactivity of gelsolin within samples and between antibodies varied greatly. In several instances multiple bands were identified corresponding to different gelsolin forms by one antibody but not identified by another. Moreover in some instances immunoreactivity depended on the source of gelsolin e.g. plasma or CSF. Additionally some smaller forms of gelsolin were identified by mass spectrometry but not by any antibody. Recombinant gelsolin was used as reference sample. Conclusions Orthogonal validation using specific monoclonal or polyclonal antibodies may reject biomarker candidates from further studies based on misleading or even false quantitation of those proteins which circulate in various forms in body fluids. Background The development of global proteomic profiling in the mid-1990 s raised the expectations for
