PMID- 37634829
OWN - NLM
STAT- MEDLINE
DCOM- 20230918
LR  - 20230918
IS  - 1873-4863 (Electronic)
IS  - 0168-1656 (Linking)
VI  - 375
DP  - 2023 Sep 20
TI  - Epitope mapping of nanobodies binding the Alzheimer's disease receptor SORLA.
PG  - 17-27
LID - S0168-1656(23)00150-5 [pii]
LID - 10.1016/j.jbiotec.2023.08.005 [doi]
AB  - Reduced levels of the Sortilin-related receptor with A-type repeats (SORLA) in 
      different brain regions as well as in the cerebrospinal fluid have been 
      associated with Alzheimer's disease. Methods and reagents to develop reliable 
      detection assays to quantify SORLA and its specific isoforms are therefore much 
      needed. Nanobodies (Nbs) are unique biomolecules derived from the blood of 
      camelids that display advantageous physicochemical and antigen affinity 
      properties, making them attractive tools with great relevance to both diagnostic 
      and therapeutic applications. Here, we purified and characterized eight Nbs that 
      were isolated from the blood of an alpaca immunized with the recombinant 
      extracellular domain of SORLA. The selected Nbs showed high affinity to SORLA in 
      the low nanomolar range as observed by surface plasmon resonance. For mapping of 
      the Nbs' epitopes within the antigen, we transiently transfected HEK293 cells 
      with a panel of SORLA deletion constructs, and developed a protocol of 
      immunostaining by applying fluorescent dye conjugated Nbs. With this method, we 
      showed that the selected Nbs specifically recognize a part of SORLA containing 
      Fibronectin-type III domains, representing promising tools not only for disease 
      clarifying research, but also for translational medicine as candidates for 
      clinical diagnostic purposes.
CI  - Copyright (c) 2023 The Authors. Published by Elsevier B.V. All rights reserved.
FAU - Monti, Giulia
AU  - Monti G
AD  - Department of Biomedicine, Aarhus University, Hoegh‑Guldbergs Gade 10, 8000 
      Aarhus C, Denmark.
FAU - Vincke, Cecile
AU  - Vincke C
AD  - Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, 
      Brussels, Belgium; Myeloid Cell Immunology Lab, VIB Center for Inflammation 
      Research, Brussels, Belgium.
FAU - Lunding, Melanie
AU  - Lunding M
AD  - Department of Biomedicine, Aarhus University, Hoegh‑Guldbergs Gade 10, 8000 
      Aarhus C, Denmark.
FAU - Jensen, Anne Mette G
AU  - Jensen AMG
AD  - Department of Biomedicine, Aarhus University, Hoegh‑Guldbergs Gade 10, 8000 
      Aarhus C, Denmark.
FAU - Madsen, Peder
AU  - Madsen P
AD  - Department of Biomedicine, Aarhus University, Hoegh‑Guldbergs Gade 10, 8000 
      Aarhus C, Denmark.
FAU - Muyldermans, Serge
AU  - Muyldermans S
AD  - Laboratory of Cellular and Molecular Immunology, Vrije Universiteit Brussel, 
      Brussels, Belgium.
FAU - Kjolby, Mads
AU  - Kjolby M
AD  - Department of Biomedicine, Aarhus University, Hoegh‑Guldbergs Gade 10, 8000 
      Aarhus C, Denmark; Department of Clinical Pharmacology and Steno Diabetes Center 
      Aarhus, Aarhus University Hospital, Denmark.
FAU - Andersen, Olav M
AU  - Andersen OM
AD  - Department of Biomedicine, Aarhus University, Hoegh‑Guldbergs Gade 10, 8000 
      Aarhus C, Denmark. Electronic address: o.andersen@biomed.au.dk.
LA  - eng
PT  - Journal Article
DEP - 20230826
PL  - Netherlands
TA  - J Biotechnol
JT  - Journal of biotechnology
JID - 8411927
RN  - 0 (Single-Domain Antibodies)
RN  - 0 (Epitopes)
SB  - IM
MH  - Humans
MH  - *Single-Domain Antibodies/genetics
MH  - Epitope Mapping
MH  - *Alzheimer Disease
MH  - HEK293 Cells
MH  - Epitopes
OTO - NOTNLM
OT  - Alzheimer's disease
OT  - Epitope mapping
OT  - Nanobodies
OT  - SORLA
COIS- Declaration of Competing Interest The authors declare the following financial 
      interests/personal relationships which may be considered as potential competing 
      interests: Olav Andersen reports financial support was provided by EU Joint 
      Programme Neurodegenerative Disease Research. Olav Andersen reports a 
      relationship with Retromer Therapeutics that includes: board membership, 
      consulting or advisory, equity or stocks, and funding grants.
EDAT- 2023/08/28 00:41
MHDA- 2023/09/18 12:44
CRDT- 2023/08/27 19:27
PHST- 2023/05/09 00:00 [received]
PHST- 2023/08/20 00:00 [revised]
PHST- 2023/08/24 00:00 [accepted]
PHST- 2023/09/18 12:44 [medline]
PHST- 2023/08/28 00:41 [pubmed]
PHST- 2023/08/27 19:27 [entrez]
AID - S0168-1656(23)00150-5 [pii]
AID - 10.1016/j.jbiotec.2023.08.005 [doi]
PST - ppublish
SO  - J Biotechnol. 2023 Sep 20;375:17-27. doi: 10.1016/j.jbiotec.2023.08.005. Epub 
      2023 Aug 26.