Characterization of Anti‐SOD1 Antibodies and Detection of Intermediary SOD1 Oligomers
Bibliographic record
Abstract
Mutations in the gene encoding the enzyme copper‐zinc superoxide dismutase (SOD1) are linked to 10–20% of familial amyotrophic lateral sclerosis (fALS) cases. The mutations lead to misfolding and self‐assembly of SOD1 into toxic oligomers and aggregates, resulting in motor neuron degeneration. However, the molecular mechanisms underlying SOD1 aggregation and toxicity are unclear. Characterization and detection of SOD1 oligomers is particularly challenging due to their metastable nature. Antibodies against oligomeric and misfolded SOD1 forms are useful tools for this purpose, provided their specificity and selectivity are well characterized, which is not always the case. In this study, we set out to characterize three novel anti‐misfolded SOD1 antibodies and compare them with two commercial anti‐misfolded SOD1 antibodies raised against G93A SOD1. As controls, we compared the reactivity of these antibodies to two polyclonal anti‐SOD1 antibodies that were expected to be insensitive to misfolding. We asked whether the antibodies could distinguish among native and misfolded conformations of SOD1 and whether any of them were specific for the elusive oligomeric forms of the protein. To follow SOD1 conformational change and aggregation kinetics, wild‐type and G93A SOD1 were incubated under reducing conditions and the reactions were followed using thioflavin T (ThT) fluorescence, electron microscopy (EM), and dot blots with each antibody. Control reactions were followed under non‐reducing conditions. The three new anti‐misfolded SOD1 and one of the commercial antibodies (B8H10) showed maximum affinity at the time oligomers were expected to dominate and lost affinity with fibril formation. Unexpectedly, the two polyclonal antibodies also showed preference for oligomers, especially of G93A SOD1. Surprisingly, the other commercial anti‐misfolded SOD1 antibody, C4F6, showed specificity for G93A SOD1 regardless of misfolding. Two of the new antibodies showed preference for early or late oligomers, suggesting that they could expand currently detectable temporal resolution. In conclusion, our data indicate that certain antibodies are highly sensitive to the misfolded form of SOD1, whereas others are insensitive to either mutation or aggregation. Support or Funding Information RGK Foundation
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.001 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.001 |
| Bibliometrics | 0.001 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.001 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 0.000 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".