title: TDP-43 protein variants as biomarkers in amyotrophic lateral sclerosis.
abstract: TDP-43 aggregates accumulate in individuals affected by amyotrophic lateral sclerosis (ALS) and other neurodegenerative diseases, representing potential diagnostic and therapeutic targets. Using an atomic force microscopy based biopanning protocol developed in our lab, we previously isolated 23 TDP-43 reactive antibody fragments with preference for human ALS brain tissue relative to frontotemporal dementia, a related neurodegeneration, and healthy samples from phage-displayed single chain antibody fragment (scFv) libraries. Here we further characterize the binding specificity of these different scFvs and identify which ones have promise for detecting ALS biomarkers in human brain tissue and plasma samples.
  
  
  
  We developed a sensitive capture ELISA for detection of different disease related TDP-43 variants using the scFvs identified from the ALS biopanning. We show that a wide variety of disease selective TDP-43 variants are present in ALS as the scFvs show different reactivity profiles amongst the ALS cases. When assaying individual human brain tissue cases, three scFvs (ALS-TDP6, ALS-TDP10 and ALS-TDP14) reacted with all the ALS cases and 12 others reacted with the majority of the ALS cases, and none of the scFvs reacted with any control samples. When assaying individual human plasma samples, 9 different scFvs reacted with all the sporadic ALS samples and again none of them reacted with any control samples. These 9 different scFvs had different patterns of reactivity with plasma samples obtained from chromosome 9 open reading frame 72 (c9orf72) cases indicating that these familial ALS genetic variants may display different TDP-43 pathology than sporadic ALS cases.
  
  
  
  These results indicated that a range of disease specific TDP-43 variants are generated in ALS patients with different variants being generated in sporadic and familial cases. We show that a small panel of scFvs recognizing different TDP-43 variants can generate a neuropathological and plasma biomarker profile with potential to distinguish different TDP-43 pathologies.
authors.affiliation: Chemical Engineering, The School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, AZ, 85287-6106, USA.
authors.forename: Stephanie M
authors.initials: SM
authors.lastname: Williams
authors.affiliation: Departments of Pathology and Neurology, Georgetown University Medical Center, Washington, DC, 20057, USA.
authors.forename: Galam
authors.initials: G
authors.lastname: Khan
authors.affiliation: Departments of Pathology and Neurology, Georgetown University Medical Center, Washington, DC, 20057, USA.
authors.forename: Brent T
authors.initials: BT
authors.lastname: Harris
authors.affiliation: Department of Neurosciences, University of California, San Diego School of Medicine, La Jolla, CA, 92093-0624, USA.
authors.forename: John
authors.initials: J
authors.lastname: Ravits
authors.affiliation: Chemical Engineering, The School for Engineering of Matter, Transport and Energy, Arizona State University, Tempe, AZ, 85287-6106, USA. sierks@asu.edu.
authors.forename: Michael R
authors.initials: MR
authors.lastname: Sierks
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000690:Amyotrophic Lateral Sclerosis / Q000097:blood / Q000235:genetics / Q000378:metabolism*; D015415:Biomarkers / Q000097:blood / Q000378:metabolism; D000073885:C9orf72 Protein; D004268:DNA-Binding Proteins / Q000097:blood / Q000276:immunology / Q000378:metabolism*; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D011506:Proteins / Q000235:genetics; D057127:Single-Chain Antibodies / Q000097:blood / Q000378:metabolism
chemical_list: D015415:Biomarkers; D000073885:C9orf72 Protein; C568651:C9orf72 protein, human; D004268:DNA-Binding Proteins; D011506:Proteins; D057127:Single-Chain Antibodies; C000598624:TARDBP protein, human
keywords: Amyotrophic lateral sclerosis; Biomarker; Brain tissue; Plasma; TDP-43 variants; scFv