Generation and characterization of antagonistic anti-human CD39 nanobodies
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Generation and characterization of antagonistic anti-human CD39 nanobodies. / Menzel, Stephan; Duan, Yinghui; Hambach, Julia; Albrecht, Birte; Wendt-Cousin, Dorte; Winzer, Riekje; Tolosa, Eva; Rissiek, Anne; Guse, Andreas H; Haag, Friedrich; Magnus, Tim; Koch-Nolte, Friedrich; Rissiek, Björn.
In: FRONT IMMUNOL, Vol. 15, 2024, p. 1328306.Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review
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TY - JOUR
T1 - Generation and characterization of antagonistic anti-human CD39 nanobodies
AU - Menzel, Stephan
AU - Duan, Yinghui
AU - Hambach, Julia
AU - Albrecht, Birte
AU - Wendt-Cousin, Dorte
AU - Winzer, Riekje
AU - Tolosa, Eva
AU - Rissiek, Anne
AU - Guse, Andreas H
AU - Haag, Friedrich
AU - Magnus, Tim
AU - Koch-Nolte, Friedrich
AU - Rissiek, Björn
N1 - Copyright © 2024 Menzel, Duan, Hambach, Albrecht, Wendt-Cousin, Winzer, Tolosa, Rissiek, Guse, Haag, Magnus, Koch-Nolte and Rissiek.
PY - 2024
Y1 - 2024
N2 - CD39 is the major enzyme controlling the levels of extracellular adenosine triphosphate (ATP) via the stepwise hydrolysis of ATP to adenosine diphosphate (ADP) and adenosine monophosphate (AMP). As extracellular ATP is a strong promoter of inflammation, monoclonal antibodies (mAbs) blocking CD39 are utilized therapeutically in the field of immune-oncology. Though anti-CD39 mAbs are highly specific for their target, they lack deep penetration into the dense tissue of solid tumors, due to their large size. To overcome this limitation, we generated and characterized nanobodies that targeted and blocked human CD39. From cDNA-immunized alpacas we selected 16 clones from seven nanobody families that bind to two distinct epitopes of human CD39. Among these, clone SB24 inhibited the enzymatic activity of CD39. Of note, SB24 blocked ATP degradation by both soluble and cell surface CD39 as a 15kD monomeric nanobody. Dimerization via fusion to an immunoglobulin Fc portion further increased the blocking potency of SB24 on CD39-transfected HEK cells. Finally, we confirmed the CD39 blocking properties of SB24 on human PBMCs. In summary, SB24 provides a new small biological antagonist of human CD39 with potential application in cancer therapy.
AB - CD39 is the major enzyme controlling the levels of extracellular adenosine triphosphate (ATP) via the stepwise hydrolysis of ATP to adenosine diphosphate (ADP) and adenosine monophosphate (AMP). As extracellular ATP is a strong promoter of inflammation, monoclonal antibodies (mAbs) blocking CD39 are utilized therapeutically in the field of immune-oncology. Though anti-CD39 mAbs are highly specific for their target, they lack deep penetration into the dense tissue of solid tumors, due to their large size. To overcome this limitation, we generated and characterized nanobodies that targeted and blocked human CD39. From cDNA-immunized alpacas we selected 16 clones from seven nanobody families that bind to two distinct epitopes of human CD39. Among these, clone SB24 inhibited the enzymatic activity of CD39. Of note, SB24 blocked ATP degradation by both soluble and cell surface CD39 as a 15kD monomeric nanobody. Dimerization via fusion to an immunoglobulin Fc portion further increased the blocking potency of SB24 on CD39-transfected HEK cells. Finally, we confirmed the CD39 blocking properties of SB24 on human PBMCs. In summary, SB24 provides a new small biological antagonist of human CD39 with potential application in cancer therapy.
KW - Humans
KW - Single-Domain Antibodies/pharmacology
KW - Adenosine Triphosphate/metabolism
KW - Adenosine Monophosphate
KW - Adenosine Diphosphate/metabolism
U2 - 10.3389/fimmu.2024.1328306
DO - 10.3389/fimmu.2024.1328306
M3 - SCORING: Journal article
C2 - 38590528
VL - 15
SP - 1328306
JO - FRONT IMMUNOL
JF - FRONT IMMUNOL
SN - 1664-3224
ER -
