期刊
BIOLOGICAL CHEMISTRY
卷 388, 期 6, 页码 651-658出版社
WALTER DE GRUYTER & CO
DOI: 10.1515/BC.2007.070
关键词
disease markers; formalin fixation; histology; immunohistochemistry; monoclonal antibodies; phage display
We describe the direct isolation of specific antibodies on formalin-fixed, paraffin-embedded (FFPE) tissue. The technique involves subtractive selection of a large and highly diverse combinatorial human antibody phage library (HuCAL (R)) on lymphocyte FFPE tissue sections. Tissue sections from normal human tonsil tissue were used to deplete the library of binders to most housekeeping proteins. Mantle-cell lymphoma tissue was used for positive selection and enrichment of mantle cell or tumor-specific antibody phage. We established a high-throughput immunohistochemical method for screening of antibody clones selected from FFPE tissue. One recombinant antibody showed specific staining for inter-follicular and mantle cells in FFPE tissue. Immunoprecipitation with this antibody and subsequent mass spectrometric analysis revealed specificity for vimentin.
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