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

Autor: Claire Perruchini, Jean-Pierre Bourgeois, François Rougeon, Pierre Lafaye, Charles Duyckaerts, Frédéric Pecorari
Přispěvatelé: Neurologie Expérimentale et Thérapeutique, IFR70-Institut National de la Santé et de la Recherche Médicale (INSERM), Unité de Biotechnologie, Biocatalyse et Biorégulation (U3B), Université de Nantes (UN)-Centre National de la Recherche Scientifique (CNRS), Génétique Humaine et Fonctions Cognitives, Institut Pasteur [Paris] (IP)-Centre National de la Recherche Scientifique (CNRS), Génétique et Biochimie du Développement, Institut Pasteur [Paris]-Centre National de la Recherche Scientifique (CNRS)
Rok vydání: 2009
Předmět:
Male
medicine.drug_class
Gene Expression
Enzyme-Linked Immunosorbent Assay
Monoclonal antibody
Pathology and Forensic Medicine
law.invention
03 medical and health sciences
Cellular and Molecular Neuroscience
Immunolabeling
0302 clinical medicine
Alzheimer Disease
law
Glial Fibrillary Acidic Protein
medicine
Animals
Humans
RNA
Messenger
Gene Library
030304 developmental biology
Cerebral Cortex
0303 health sciences
Messenger RNA
Glial fibrillary acidic protein
biology
Brain Neoplasms
Antibodies
Monoclonal
[SDV.IMM.IMM]Life Sciences [q-bio]/Immunology/Immunotherapy
Molecular biology
Recombinant Proteins
3. Good health
Molecular Weight
Ribosome display
biology.protein
Recombinant DNA
Immunohistochemistry
Neurology (clinical)
Antibody
Immunoglobulin Heavy Chains
Camelids
New World
030217 neurology & neurosurgery
Single-Chain Antibodies
Zdroj: Acta Neuropathologica
Acta Neuropathologica, 2009, 118 (5), pp.685-95. ⟨10.1007/s00401-009-0572-6⟩
Acta Neuropathologica, Springer Verlag, 2009, 118 (5), pp.685-95. ⟨10.1007/s00401-009-0572-6⟩
ISSN: 1432-0533
0001-6322
DOI: 10.1007/s00401-009-0572-6
Popis: International audience; Camelids produce antibodies made of homodimeric heavy chains, and the antigen-binding region being composed of a single domain called V(H)H. These V(H)Hs 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(H)Hs, 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(H)H-coding sequences were selectively amplified. Three V(H)Hs with a high affinity for GFAP and their corresponding mRNA were selected by ribosome display. Large quantities of the recombinant V(H)Hs 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(H)H 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.
Databáze: OpenAIRE
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