Selection of vimentin-specific antibodies from the HuCAL® phage display library by subtractive panning on formalin-fixed, paraffin-embedded tissue

The antigen retrieval (AR) technique, which is predominantly based on high-temperature heating of tissues, is used as a non-enzymatic pretreatment for immunohistochemical staining of formalin-fixed, paraffin-embedded tissue sections. It has been widely applied in pathology and analytical morphology. The existence of a growing body of literature on the AR technique raises a number of interesting issues for the further development of AR. These issues include the use of a “test battery” and the concept of “maximal retrieval” applied to the selection of optimal test protocols for the standardization of AR. (J Histochem Cytochem 45:327–343, 1997)

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Antibody Identification for Antigen Detection in Formalin-Fixed Paraffin-Embedded Tissue Using Phage Display and Naïve Libraries

Antibodies ◽

10.3390/antib10010004 ◽

2021 ◽

Vol 10 (1) ◽

pp. 4

Author(s):

Célestine Mairaville ◽

Pierre Martineau

Keyword(s):

Phage Display ◽

Formalin Fixation ◽

Antigen Retrieval ◽

Alternative Strategies ◽

Tissue Sections ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Antibody Libraries ◽

Formalin Fixed

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.

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Selection Strategies for Tissue-Specific Antibody Identification Using Phage Display and Naïve Libraries

10.20944/preprints202010.0607.v1 ◽

2020 ◽

Author(s):

Célestine Mairaville ◽

Pierre Martineau

Keyword(s):

Phage Display ◽

Antigen Retrieval ◽

Alternative Strategies ◽

Tissue Sections ◽

Selection Strategies ◽

Formalin Fixed Paraffin ◽

Formalin Fixed Paraffin Embedded ◽

Antibody Libraries ◽

Formalin Fixed

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.

Download Full-text