Journal: bioRxiv

Article Title: Antibody treatment targeting nitrated alpha-synuclein counteracts protein spreading pathology

doi: 10.64898/2026.05.21.726933

Figure Lengend Snippet: (A) Binding of 6G6 to biotinylated HαSyn 50-mer peptides nitrated at Tyr39 (nTyr39-50mer), Tyr125 (nTyr125-50mer), Tyr133 (nTyr133-50mer) or Tyr136 (nTyr136-50mer), as measured by electrochemiluminescent ELISA. Non-nitrated Tyr39 peptide (Tyr39-50mer) and unmodified full-length HαSyn (FL-HαSyn) were included as controls. (B) Binding of 6G6 to chemically nitrated full-length human α-synuclein (N-HαSyn), recombinant site-specifically Tyr39-nitrated full-length HαSyn (nTyr39 rec-HαSyn), or peroxynitrite-modified bovine serum albumin (N-BSA). (C) Western blot analysis showing the reactivity of 6G6, anti-3-nitrotyrosine, and total α-synuclein antibody (BD610787 αSyn) toward unmodified HαSyn, chemically nitrated HαSyn, recombinant site-specifically Tyr39-nitrated HαSyn, unmodified MsαSyn, and chemically nitrated MsαSyn.

Article Snippet: The αSyn peptides and full-length human αSyn (Abclonal) were used for ELISA, affinity, and specificity studies.

Techniques: Binding Assay, Enzyme-linked Immunosorbent Assay, Recombinant, Modification, Western Blot

Journal: bioRxiv

Article Title: Antibody treatment targeting nitrated alpha-synuclein counteracts protein spreading pathology

doi: 10.64898/2026.05.21.726933

Figure Lengend Snippet:

Article Snippet: The αSyn peptides and full-length human αSyn (Abclonal) were used for ELISA, affinity, and specificity studies.

Techniques: Binding Assay

Journal: bioRxiv

Article Title: Antibody treatment targeting nitrated alpha-synuclein counteracts protein spreading pathology

doi: 10.64898/2026.05.21.726933

Figure Lengend Snippet: (A) Experimental design. Homozygous M83 mice received a single unilateral intrastriatal injection of HαSyn PFFs or monomer on day 0. Antibody treatment with control IgG or 6G6 was started 2 h before inoculation and continued once weekly for 7 weeks. Animals were sacrificed at study week 7, 2 days after the final antibody injection. Coronal brain sections were immunostained with a specific antibody against P-αSyn. (B) Representative images from coronal sections immunostained for P-αSyn, showing pathology in the striatum, cingulate cortex, frontal cortex (FC), basolateral amygdala (Amy), substantia nigra pars compacta (SNc), substantia nigra pars reticulata (SNr), and thalamus ipsilateral to the injection side, as well as in the SNc, SNr and thalamus contralateral to the injection side. Scale bar = 50 µm. (C, D) Low-magnification images showing the regions of interest used for P-αSyn quantification in the substantia nigra and thalamus. Scale bar = 1 mm. (E) Quantification of P-αSyn-positive objects per analysed area in the substantia nigra ipsilateral and contralateral to the injection side in IgG-treated (n = 9) and 6G6-treated (n = 8) mice. Box-and-whisker plots show the median, upper and lower quartiles, and maximum and minimum values as whiskers. P < 0.05; ** P < 0.001, unpaired t -test. (F) Quantification of P-αSyn-positive objects per analysed area in the thalamus ipsilateral and contralateral to the injection side in IgG-treated (n = 9) and 6G6-treated (n = 8) mice. Box-and-whisker plots show the median, upper and lower quartiles, and maximum and minimum values as whiskers. ** P < 0.001, **** P < 0.0001; unpaired t -test.

Article Snippet: The αSyn peptides and full-length human αSyn (Abclonal) were used for ELISA, affinity, and specificity studies.

Techniques: Injection, Control, Whisker Assay

Journal: bioRxiv

Article Title: Antibody treatment targeting nitrated alpha-synuclein counteracts protein spreading pathology

doi: 10.64898/2026.05.21.726933

Figure Lengend Snippet: (A) Total protein concentrations in cerebrospinal fluid samples from healthy (n=5) controls and Parkinson’s disease (n=10) patients. (B) Total αSyn concentrations in the same CSF samples. (C) Tyr39-nitrated αSyn concentrations measured by SIMOA in CSF samples from healthy controls and Parkinson’s disease patients. Box-and-whisker plots show the median, upper and lower quartiles, and maximum and minimum values as whiskers. *** P < 0.001, unpaired t -test.

Article Snippet: The αSyn peptides and full-length human αSyn (Abclonal) were used for ELISA, affinity, and specificity studies.

Techniques: Whisker Assay

Journal: Journal of the American Chemical Society

Article Title: Time-Resolved Native Mass Spectrometry for Direct Measurement of Biomolecular Kinetics

doi: 10.1021/jacs.5c21842

Figure Lengend Snippet: Capturing an array of mixing times using theta emitters. (A) Mass spectrum of 0.8 μM green fluorescent protein (GFP) binding 2 μM Nb15 (15) and 4 μM NbAS (AS) at equilibrium. (B) Extracted mass chromatogram of apo, Nb15, and NbAS-bound GFP over three ESI power cycles acquired in one data acquisition. (C–E) Mass spectra 0.8 μM GFP binding 2 μM Nb15 and 4 μM NbAS in a theta emitter shortly after ESI voltage was turned on (C), and after (D) 1 or (E) 1.5 min of sustained voltage. The concentrations listed account for a 2-fold dilution within the Taylor cone during rapid mixing. Deconvoluted spectra are shown in Figure S9 .

Article Snippet: Briefly, codon-optimized genes for anti-alpha synuclein (NbAS), anti-GFP (Nb15), and anti-ALFA (NbALFA) were obtained from Twist Biosciences.

Techniques: Binding Assay

Journal: Journal of the American Chemical Society

Article Title: Time-Resolved Native Mass Spectrometry for Direct Measurement of Biomolecular Kinetics

doi: 10.1021/jacs.5c21842

Figure Lengend Snippet: Capturing kinetics of protein–protein interactions using theta emitters. (A, B) Biolayer interferometry (BLI) sensorgrams showing GFP binding to biotinylated (A) NbAS and (B) Nb15, with corresponding global fits (solid lines) based on a 1:1 binding model. (C) Crystal structure of Nb15 bound to GFP ( Table S1 and Figure S14 ). (D) Time-resolved native MS kinetic curves showing GFP binding to Nb15, timed using NbAS. Data from five bursts are plotted in distinct colors, fit to a 1:1 global kinetic model in which each burst is assigned its own mixing factor (α). The colored traces represent individual bursts, and solid lines denote the corresponding model fits, with each α value labeled in the matching color. (E) Kinetic rate constants and equilibrium dissociation constants ( K D ) for the GFP–Nb15 interaction timed with GFP-NbALFA and SOS–NbALFA, and for the GFP–NbAS interaction timed with GFP-NbALFA, as determined by BLI and time-resolved native mass spectrometry. A table of kinetic values for each system is provided in Table S2 .

Article Snippet: Briefly, codon-optimized genes for anti-alpha synuclein (NbAS), anti-GFP (Nb15), and anti-ALFA (NbALFA) were obtained from Twist Biosciences.

Techniques: Protein-Protein interactions, Binding Assay, Labeling, Mass Spectrometry