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2D NMR structural comparison and principal component analysis of the mAb1 aglycosylation series. (A) Spectral overlay of the 2D NMR fingerprints of mAb1-WT and mAb2 illustrating structural differences arising from primary sequence variation. (B) PCA scores plot of processed 2D NMR spectra including the mAb1 co-mix series (0–100% NGHC) and mAb2. PCA was performed using the BioHOS workflow implemented in <t>Mnova</t> <t>(mestrelab</t> research) following spectral normalization, binning, and mean-centering as described in the methods section. The first two principal components capture the majority of spectral variance, with mAb2 clearly separated from the mAb1 samples. (C) Spectral overlay of mAb1-WT and mAb1-N297G (100% aglycosylated), illustrating spectral differences associated with complete loss of Fc N-linked glycosylation. (D) PCA scores plot of the mAb1 co-mix series as a function of increasing NGHC content. Individual points correspond to single spectra from each sample. Spectra containing up to ~25% NGHC occupy a similar region of PCA space, whereas higher NGHC levels progressively diverge along the principal component axes.
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2D NMR structural comparison and principal component analysis of the mAb1 aglycosylation series. (A) Spectral overlay of the 2D NMR fingerprints of mAb1-WT and mAb2 illustrating structural differences arising from primary sequence variation. (B) PCA scores plot of processed 2D NMR spectra including the mAb1 co-mix series (0–100% NGHC) and mAb2. PCA was performed using the BioHOS workflow implemented in Mnova (mestrelab research) following spectral normalization, binning, and mean-centering as described in the methods section. The first two principal components capture the majority of spectral variance, with mAb2 clearly separated from the mAb1 samples. (C) Spectral overlay of mAb1-WT and mAb1-N297G (100% aglycosylated), illustrating spectral differences associated with complete loss of Fc N-linked glycosylation. (D) PCA scores plot of the mAb1 co-mix series as a function of increasing NGHC content. Individual points correspond to single spectra from each sample. Spectra containing up to ~25% NGHC occupy a similar region of PCA space, whereas higher NGHC levels progressively diverge along the principal component axes.

Journal: mAbs

Article Title: 2D NMR assessment of structural consistency and functional relationships in monoclonal antibodies

doi: 10.1080/19420862.2026.2672774

Figure Lengend Snippet: 2D NMR structural comparison and principal component analysis of the mAb1 aglycosylation series. (A) Spectral overlay of the 2D NMR fingerprints of mAb1-WT and mAb2 illustrating structural differences arising from primary sequence variation. (B) PCA scores plot of processed 2D NMR spectra including the mAb1 co-mix series (0–100% NGHC) and mAb2. PCA was performed using the BioHOS workflow implemented in Mnova (mestrelab research) following spectral normalization, binning, and mean-centering as described in the methods section. The first two principal components capture the majority of spectral variance, with mAb2 clearly separated from the mAb1 samples. (C) Spectral overlay of mAb1-WT and mAb1-N297G (100% aglycosylated), illustrating spectral differences associated with complete loss of Fc N-linked glycosylation. (D) PCA scores plot of the mAb1 co-mix series as a function of increasing NGHC content. Individual points correspond to single spectra from each sample. Spectra containing up to ~25% NGHC occupy a similar region of PCA space, whereas higher NGHC levels progressively diverge along the principal component axes.

Article Snippet: Easy Comparability of Higher Order Structure (ECHOS) analysis was performed using the Mnova software package developed by Mestrelab Research.

Techniques: Comparison, Sequencing, Glycoproteomics