Novel SOD1 monoclonal antibodies against the electrostatic loop preferentially detect misfolded SOD1 aggregates

Yuxing Xia; Zhijuan Chen; Guilian Xu; David R Borchelt; Jacob I Ayers; Benoit I Giasson

doi:10.1016/j.neulet.2020.135553

Novel SOD1 monoclonal antibodies against the electrostatic loop preferentially detect misfolded SOD1 aggregates

Neurosci Lett. 2021 Jan 18:742:135553. doi: 10.1016/j.neulet.2020.135553. Epub 2020 Dec 17.

Authors

Yuxing Xia¹, Zhijuan Chen¹, Guilian Xu¹, David R Borchelt², Jacob I Ayers³, Benoit I Giasson⁴

Affiliations

¹ Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
² Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA; McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA.
³ Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA; McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Institute for Neurodegenerative Diseases, Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, CA, 94158, USA; Department of Neurology, Weill Institute for Neurosciences, University of California, San Francisco, San Francisco, CA, 94158, USA.
⁴ Department of Neuroscience, College of Medicine, University of Florida, Gainesville, FL 32610, USA; Center for Translational Research in Neurodegenerative Disease, College of Medicine, University of Florida, Gainesville, FL 32610, USA; McKnight Brain Institute, College of Medicine, University of Florida, Gainesville, FL 32610, USA. Electronic address: bgiasson@ufl.edu.

Abstract

Amyotrophic lateral sclerosis (ALS) is a progressive neurological disease that leads to motor neuron degeneration and paralysis. Superoxide dismutase (SOD1) mutations are the second most common cause of familial ALS and are responsible for up to 20 % of familial ALS cases. In ALS patients, SOD1 can form toxic misfolded aggregates that deposit in the brain and spinal cord. To better detect SOD1 aggregates and expand the repertoire of conformational SOD1 antibodies, SOD1 monoclonal antibodies were generated by immunizing SOD1 knockout mice with an SOD1 fragment consisting of amino acids 129-146, which make up part of the electrostatic loop. A series of hybridomas secreting antibodies were screened and five different SOD1 monoclonal antibodies (2C10, 2F8, 4B11, 5H5, and 5A10) were found to preferentially detect denatured or aggregated SOD1 by enzyme-linked immunosorbent assay (ELISA), filter trap assay, and immunohistochemical analysis in SOD1 mouse models. The staining with these antibodies was compared to Campbell-Switzer argyrophilic reactivity of pathological inclusions. These new conformational selective SOD1 antibodies will be useful for clinical diagnosis of SOD1 ALS and potentially for passive immunotherapy.

Keywords: Amyotrophic lateral sclerosis; Antibodies; Frontotemporal dementia; Pathology; Silver stain; Superoxide dismutase 1.

Publication types

Research Support, N.I.H., Extramural

MeSH terms

Amino Acid Sequence
Animals
Antibodies, Monoclonal / chemistry
Antibodies, Monoclonal / metabolism*
HEK293 Cells
Humans
Mice
Mice, Inbred BALB C
Mice, Knockout
Protein Aggregates / physiology*
Protein Folding*
Static Electricity*
Superoxide Dismutase-1 / chemistry
Superoxide Dismutase-1 / metabolism*

Substances

Antibodies, Monoclonal
Protein Aggregates
SOD1 protein, human
Superoxide Dismutase-1

Abstract

Publication types

MeSH terms

Substances

Grants and funding