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01-11-2009 | Original Paper

Llama V_HH antibody fragments against GFAP: better diffusion in fixed tissues than classical monoclonal antibodies

Authors: Claire Perruchini, Frederic Pecorari, Jean-Pierre Bourgeois, Charles Duyckaerts, François Rougeon, Pierre Lafaye

Published in: Acta Neuropathologica | Issue 5/2009

Abstract

Camelids produce antibodies made of homodimeric heavy chains, and the antigen-binding region being composed of a single domain called V_HH. These V_HHs are much smaller than complete IgG. They are also more thermostable and more soluble in water; they should, therefore, diffuse more readily in the tissues. V_HHs, expressed in bacteria, are easier to produce than conventional monoclonal antibodies. Because of these special characteristics, these antibody fragments could have interesting developments in immunohistochemistry and in the development of biomarkers. To test the possibility of their use in immunohistochemistry (IHC), we selected the glial fibrillary acidic protein (GFAP), a well-known marker of astrocytes. One alpaca (Lama pacos) was immunized against GFAP. Lymphocytes were isolated; the DNA was extracted; the V_HH-coding sequences were selectively amplified. Three V_HHs with a high affinity for GFAP and their corresponding mRNA were selected by ribosome display. Large quantities of the recombinant V_HHs coupled with different tags were harvested from transfected bacteria. One of them was shown to immunolabel strongly and specifically to GFAP of human astrocytes in tissue sections. The quality of the IHC was comparable or, in some aspects, superior to the quality obtained with conventional IgG. The V_HH was shown to diffuse on a longer distance than conventional monoclonal antibodies in fixed cortical tissue: a property that may be useful in immunolabeling of thick sections.

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Title: Llama VHH antibody fragments against GFAP: better diffusion in fixed tissues than classical monoclonal antibodies
Authors: Claire Perruchini
Frederic Pecorari
Jean-Pierre Bourgeois
Charles Duyckaerts
François Rougeon
Pierre Lafaye
Publication date: 01-11-2009
Publisher: Springer-Verlag
Published in: Acta Neuropathologica / Issue 5/2009
Print ISSN: 0001-6322
Electronic ISSN: 1432-0533
DOI: https://doi.org/10.1007/s00401-009-0572-6

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Llama V_HH antibody fragments against GFAP: better diffusion in fixed tissues than classical monoclonal antibodies

Abstract

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Abstract

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