Nicole A. Rivera-Fuentes, Addison E. Bergman, Brandon T. Ruotolo. “Conjugation location-dependent stability shifts in antibody drug conjugate mimics revealed by collision induced unfolding.” ChemRxiv (). https://doi.org/10.26434/chemrxiv.15007469/v1

The main paper refers to supporting information, but no separate supporting information file was found among the files available from the ChemRxiv record

Deglycosylated Rituximab
Native-like charge states 26+ – 23+
Deconvoluted mass 144,225 ± 2 Da
TWΩN2 (source temperatures 25 °C and 40 °C) 71.0 ± 0.3 nm²
Main, pp. 5–6
Fc-conjugated Rituximab
Deconvoluted mass DAR 1: 144,666 ± 1 Da; DAR 2: 145,137.5 ± 0.7 Da
Average DAR 1.7
Low-intensity DAR 3 and DAR 4 ions 2 – 8 %
Remaining unconjugated antibody ~6 %
TWΩN2 (24+) DAR 1: 71.8 ± 0.4 nm²; DAR 2: 72.1 ± 0.4 nm²
CIU50-2 (24+) DAR 0: 105 ± 1 V; DAR 1: 109.6 ± 0.2 V; DAR 2: 112 ± 1 V
CIU50-2 gas-phase stability increase DAR 0 to DAR 1: 4.4 %; DAR 0 to DAR 2: 6.7 %
Main, pp. 5–7
Fab-conjugated Rituximab
Deconvoluted mass DAR 0: 144,225 ± 2 Da; DAR 1: 144,824 ± 1 Da; DAR 2: 145,424 ± 3 Da; DAR 3: 146,023 ± 1 Da; DAR 4: 146,622 ± 1 Da
Average DAR 1.3
TWΩN2 (24+) DAR 0: 71.0 ± 0.3 nm²; DAR 1: 72.5 ± 0.4 nm²; DAR 2: 74.0 ± 0.4 nm²
CIU50-1 (24+) DAR 0: 66.59 ± 0.18 V; DAR 1: 71.55 ± 0.19 V; DAR 2: 74.6 ± V
CIU50-1 gas-phase stability increase DAR 0 to DAR 1: 7.2 %; DAR 0 to DAR 2: 11.5 %
Main, pp. 5–7
Glycosylated Rituximab Fab fragment
Charge states and mass 15+ to 12+; 47,036 ± 4 Da
Main, p. 6
Glycosylated Rituximab Fc fragment
Charge states and mass 12+ to 11+; 53,258 ± 21 Da
Main, p. 6
Fab-conjugated Rituximab Fab fragment
Mass 47,629 ± 4 Da and 48,229.9 ± 0.1 Da
CIU50 transition increase CIU50-1: ~17 – 22 %; CIU50-2: 12 – 19 %; CIU50-3: 2 – 9 %
Main, pp. 6, 8
Deglycosylated Rituximab Fc fragment
Charge states and mass 11+ to 10+; 50,280 ± 16 Da
Main, p. 6
Fc-conjugated Rituximab Fc fragment
Fc-conjugated ion populations 50,277 ± 22 Da, 50,712 ± 13 Da, and 51,228 ± 21 Da
CIU50-3 DAR 0: 84 ± 4 V; DAR 1: 123 ± 4 V; ~37 % increase