TY - JOUR
T1 - Employing nanobodies for immune landscape profiling by PET imaging in mice
AU - Teunissen, Abraham J. P.
AU - Abousaway, Omar B.
AU - Munitz, Jazz
AU - van Leent, Mandy M. T.
AU - Toner, Yohana C.
AU - Priem, Bram
AU - Senders, Max L.
AU - Pérez-Medina, Carlos
AU - Mulder, Willem J. M.
AU - Rashidian, Mohammad
N1 - Funding Information: This work was supported by National Institutes of Health (NIH) grants R01 CA220234 , R01 HL144072 , and P01 HL131478 , NWO /ZonMW Vici 91818622 (all to W.J.M.M.), NIH 1K22CA226040 (to M.R.), and the Dana-Farber Cancer Institute Innovation Research Fund Basic Research Grant (to M.R). B.P. was supported by the AMC Ph.D. Scholarship. The anti-CD11b nanobody and sortase 7M plasmids were kind gifts from Dr. Hidde Ploegh’s laboratory. Publisher Copyright: © 2021 The Authors
PY - 2021/6/18
Y1 - 2021/6/18
N2 - Noninvasive immunoimaging holds great potential for studying and stratifying disease as well as therapeutic efficacy. Radiolabeled single-domain antibody fragments (i.e., nanobodies) are appealing probes for immune landscape profiling, as they display high stability, rapid targeting, and excellent specificity, while allowing extremely sensitive nuclear readouts. Here, we present a protocol for radiolabeling an anti-CD11b nanobody and studying its uptake in mice by a combination of positron emission tomography imaging, ex vivo gamma counting, and autoradiography. Our protocol is applicable to nanobodies against other antigens. For complete details on the use and execution of this protocol, please see Priem et al. (2020), Senders et al. (2019), or Rashidian et al. (2017).
AB - Noninvasive immunoimaging holds great potential for studying and stratifying disease as well as therapeutic efficacy. Radiolabeled single-domain antibody fragments (i.e., nanobodies) are appealing probes for immune landscape profiling, as they display high stability, rapid targeting, and excellent specificity, while allowing extremely sensitive nuclear readouts. Here, we present a protocol for radiolabeling an anti-CD11b nanobody and studying its uptake in mice by a combination of positron emission tomography imaging, ex vivo gamma counting, and autoradiography. Our protocol is applicable to nanobodies against other antigens. For complete details on the use and execution of this protocol, please see Priem et al. (2020), Senders et al. (2019), or Rashidian et al. (2017).
KW - Antibody
KW - Immunology
KW - Model Organisms
KW - Molecular Biology
KW - Molecular/Chemical Probes
UR - http://www.scopus.com/inward/record.url?scp=85103729243&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.xpro.2021.100434
DO - https://doi.org/10.1016/j.xpro.2021.100434
M3 - Article
C2 - 33899016
SN - 2666-1667
VL - 2
JO - STAR Protocols
JF - STAR Protocols
IS - 2
M1 - 100434
ER -