Development of Antibody and Nanobody Tools for P2X7
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Development of Antibody and Nanobody Tools for P2X7. / Stähler, Tobias; Danquah, Welbeck; Demeules, Melanie; Gondé, Henri; Hardet, Romain; Haag, Friedrich; Adriouch, Sahil; Koch-Nolte, Friedrich; Menzel, Stephan.
The P2X7 Receptor: Methods and Protocols. Hrsg. / Annette Nicke. 1. Aufl. New York, NY : HUMANA PRESS INC, 2022. S. 99-127 (Methods Mol Biol; Band 2510).Publikationen: SCORING: Beitrag in Buch/Sammelwerk › SCORING: Beitrag in Sammelwerk › Forschung › Begutachtung
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TY - CHAP
T1 - Development of Antibody and Nanobody Tools for P2X7
AU - Stähler, Tobias
AU - Danquah, Welbeck
AU - Demeules, Melanie
AU - Gondé, Henri
AU - Hardet, Romain
AU - Haag, Friedrich
AU - Adriouch, Sahil
AU - Koch-Nolte, Friedrich
AU - Menzel, Stephan
N1 - © 2022. The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature.
PY - 2022
Y1 - 2022
N2 - Antibodies that recognize the ATP-gated P2X7 ion channel are etablished research tools. Nanobodies correspond to the antigen-binding variable immunoglobulin domain (VHH) of heavy chain antibodies that naturally occur in camelids. Nanobodies display better solubility than the variable domains (VH) of conventional antibodies. Therefore, it is much easier to construct bivalent and multivalent fusion proteins with nanobodies than with VH domains or with paired VH-VL domains. Moreover, nanobodies can bind functional crevices that are poorly accessbile to conventional VH-VL domains. This makes nanobodies particulary well suited as functional modulators. Here we provide protocols to raise antibodies and nanobodies against mouse and human P2X7 using cDNA-immunization. This approach evokes antibodies and nanobodies that recognize the P2X7 ion channel in native confirmation, some of which inhibit or potentiate gating of P2X7 by extracellular ATP. Furthermore, we developed protocols for producing P2X7-specific nanobodies and antibodies in vivo using rAAV vectors (AAVnano). This approach can be used either to durably inhibit or potentiate P2X7 function in vivo, or to deplete P2X7-expressing cells.
AB - Antibodies that recognize the ATP-gated P2X7 ion channel are etablished research tools. Nanobodies correspond to the antigen-binding variable immunoglobulin domain (VHH) of heavy chain antibodies that naturally occur in camelids. Nanobodies display better solubility than the variable domains (VH) of conventional antibodies. Therefore, it is much easier to construct bivalent and multivalent fusion proteins with nanobodies than with VH domains or with paired VH-VL domains. Moreover, nanobodies can bind functional crevices that are poorly accessbile to conventional VH-VL domains. This makes nanobodies particulary well suited as functional modulators. Here we provide protocols to raise antibodies and nanobodies against mouse and human P2X7 using cDNA-immunization. This approach evokes antibodies and nanobodies that recognize the P2X7 ion channel in native confirmation, some of which inhibit or potentiate gating of P2X7 by extracellular ATP. Furthermore, we developed protocols for producing P2X7-specific nanobodies and antibodies in vivo using rAAV vectors (AAVnano). This approach can be used either to durably inhibit or potentiate P2X7 function in vivo, or to deplete P2X7-expressing cells.
KW - Adenosine Triphosphate
KW - Animals
KW - Antibodies
KW - Immunoglobulin Heavy Chains
KW - Mice
KW - Single-Domain Antibodies/chemistry
U2 - 10.1007/978-1-0716-2384-8_6
DO - 10.1007/978-1-0716-2384-8_6
M3 - SCORING: Contribution to collected editions/anthologies
C2 - 35776322
SN - 978-1-0716-2383-1
T3 - Methods Mol Biol
SP - 99
EP - 127
BT - The P2X7 Receptor
A2 - Nicke, Annette
PB - HUMANA PRESS INC
CY - New York, NY
ER -
