Abstract
Immunohistochemistry is a widely used technique for research and diagnostic purposes that relies on the recognition by antibodies of antigens expressed in tissues. However, tissue processing and particularly formalin fixation affect the conformation of these antigens through the formation of methylene bridges. Although antigen retrieval techniques can partially restore antigen immunoreactivity, it is difficult to identify antibodies that can recognize their target especially in formalin-fixed paraffin-embedded tissues. Most of the antibodies currently used in immunohistochemistry have been obtained by animal immunization; however, in vitro display techniques represent alternative strategies that have not been fully explored yet. This review provides an overview of phage display-based antibody selections using naïve antibody libraries on various supports (fixed cells, dissociated tissues, tissue fragments, and tissue sections) that have led to the identification of antibodies suitable for immunohistochemistry.
Highlights
Detection in Formalin-FixedImmunohistochemistry (IHC) is a procedure routinely used for research and diagnostic purposes [1] that relies on the detection by antibodies of antigens in tissue sections.Depending on the revelation system, tissue staining is monitored by bright-field or fluorescence microscopy [2], the latter being referred to as immunofluorescence (IF)
Tissue processing strongly affects the reactivity of antibodies, and the same antibody is rarely efficient on both types of tissue sections [3,4]
For an illustration of the method, the reader can refer to Figure 1 in Lykkemark et al [64]. This method allows the elution of several areas on a given slide, and prevents the loss of relevant phages fixed on regions of interest (ROI) that would not be protected by a single shadow stick
Summary
Immunohistochemistry (IHC) is a procedure routinely used for research and diagnostic purposes [1] that relies on the detection by antibodies of antigens in tissue sections. Several publications have described the use of phage-displayed peptide libraries, panned on fresh tissues, dissociated or not [32,33,34,35,36,37], and on tissue sections [38,39,40], eventually associated with laser capture microdissection. In these publications, the peptides were rarely validated by IHC or IF. Some of the illustrations were obtained from Servier Medical Art library
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