Aptamer-based enrichment of TDP-43 from human cells and tissues with quantification by HPLC-MS/MS

Pobran, Taylor D.; Yang, David; DeMarco, Mari L.

doi:10.1016/j.jneumeth.2021.109344

Cited by 5 publications

(3 citation statements)

References 42 publications

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“…For instance, knowledge of the structural and physiochemical properties of a target analyte can be leveraged to design enrichment strategies and optimize analytical procedures for large gains in analytical sensitivity. Analytical sensitivity along with molecular selectivity, based on our experience in developing TDP‐43 proteoform assays, 21 , 23 will be critical to these translational efforts. Thus, these new findings provide additional molecular‐level evidence needed to advance a much sought after antemortem, pathology‐specific biomarker for FTD with TDP‐43 pathology.…”

Section: Discussionmentioning

confidence: 99%

“…Quantification of TDP‐43 peptides was performed using our previously developed targeted high performance liquid chromatography‐mass spectrometry (LC‐MS/MS) method, 21 , 23 adapted for μLC‐MS/MS for increased analytical sensitivity and decreased sample volume requirements (see Supplemental Materials and Figure S1 ). In the MRM method, one quantifier and one qualifier ion are monitored per peptide, and quantification is performed via a single‐point internal calibrator via the addition of isotopically labeled internal standards (IS) of TDP‐43 peptides to each sample (Tables S2 and S3 ).…”

Section: Methodsmentioning

confidence: 99%

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Truncated TDP‐43 proteoforms diagnostic of frontotemporal dementia with TDP‐43 pathology

Forgrave

Moon

Hamden

et al. 2023

Alzheimer's & Dementia

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INTRODUCTIONBiomarkers of TDP‐43 pathology are needed to distinguish frontotemporal lobar degeneration with TDP‐43 pathology (FTLD‐TDP) from phenotypically related disorders. While normal physiological TDP‐43 is not a promising biomarker, low‐resolution techniques have suggested truncated forms of TDP‐43 may be specific to TDP‐43 pathology. To advance biomarker efforts for FTLD‐TDP, we employed a high‐resolution structural technique to characterize TDP‐43 post‐translational modifications in FTLD‐TDP.METHODSHigh‐resolution mass spectrometry was used to characterize TDP‐43 proteoforms in brain tissue from FTLD‐TDP, non‐TDP‐43 dementias and neuropathologically unaffected cases. Findings were then verified in a larger cohort of FTLD‐TDP and non‐TDP‐43 dementias via targeted quantitative mass spectrometry.RESULTSIn the discovery phase, truncated TDP‐43 identified FTLD‐TDP with 85% sensitivity and 100% specificity. The verification phase revealed similar findings, with 83% sensitivity and 89% specificity.DISCUSSIONThe concentration of truncated TDP‐43 proteoforms—in particular, in vivo generated C‐terminal fragments—have high diagnostic accuracy for FTLD‐TDP.HIGHLIGHTS Discovery: Truncated TDP‐43 differentiates FTLD‐TDP from related dementias. Verification: Truncated TDP‐43 concentration has high accuracy for FTLD‐TDP. TDP‐43 proteoforms <28 kDa have highest discriminatory power for TDP‐43 pathology.

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Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Truncated TDP‐43 proteoforms diagnostic of frontotemporal dementia with TDP‐43 pathology

Forgrave

Moon

Hamden

et al. 2023

Alzheimer's & Dementia

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“…8,9 Current detection methods for TDP-43 protein include enzymelinked immunosorbent assay (ELISA), western blot, atomic force microscopy-based biopanning, high-performance liquid chromatography, mass spectrometry, and surface plasmon resonance. [10][11][12][13] Given the clinical demonstrations of TDP-43 in biological fluids, there is a need for novel biosensors. Such sensors would allow for monitoring disease progression, differential diagnosis, drug screening and treatment monitoring.…”

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confidence: 99%

Antibody-Based Electrochemical Sensor for Detection of the Full-Length Phosphorylated TDP-43 Protein Biomarker of Amyotrophic Lateral Sclerosis

Wallace

Tabobondung

Esposto

et al. 2023

J. Electrochem. Soc.

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Transactive response DNA binding protein (TDP-43) is a biomarker associated with neurodegenerative diseases, specifically amyotrophic lateral sclerosis (ALS). ALS remains without treatment or a cure, and diagnosis relies on the onset of symptoms. Hence, novel methods are needed for the early detection of TDP-43 as an ALS biomarker. Toward this aim, the detection of full-length phosphorylated TDP-43 (pTDP-43) was achieved by using the electrochemical impedance spectroscopy (EIS)-based biosensor. The TDP-43 antibodies (Abs) on gold (Au) surfaces (Ab-Au) were employed as recognition probes for the protein detection. EIS was used to characterize the Ab-Au surface before and after pTDP-43 binding. In the presence of a solution redox probe, [Fe(CN)6]3-/4-, the dramatic changes in the charge-transfer resistance (Rct) values were observed after the pTDP-43 binding and were directly related to the amount of protein present in solution. Sensitivity for pTDP-43 was highly dependent on the antibody used as a recognition probe, and the limit of detection was 11 ± 6 nM with a large dynamic range, and an excellent selectivity against the common protein. The electrochemical immunosensor may be easily extended for the detection of other degenerative disease biomarkers.

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Advancing mechanistic understanding and biomarker development in amyotrophic lateral sclerosis

Thompson

Oeckl

Feneberg

et al. 2021

Expert Review of Proteomics

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Aptamer-based enrichment of TDP-43 from human cells and tissues with quantification by HPLC-MS/MS

Cited by 5 publications

References 42 publications

Truncated TDP‐43 proteoforms diagnostic of frontotemporal dementia with TDP‐43 pathology

Truncated TDP‐43 proteoforms diagnostic of frontotemporal dementia with TDP‐43 pathology

Antibody-Based Electrochemical Sensor for Detection of the Full-Length Phosphorylated TDP-43 Protein Biomarker of Amyotrophic Lateral Sclerosis

Advancing mechanistic understanding and biomarker development in amyotrophic lateral sclerosis

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