Hyphenation of strong cation exchange chromatography to native mass spectrometry for high throughput online characterization of charge heterogeneity of therapeutic monoclonal antibodies

Figures & data

Figure 1. Optimization of starting pH (a), initial gradient (b), flow rate (c), source condition (d) to improve low abundant species detection

Figure 2. Charge variants separation of seven mAbs (different pI) using the SCX-MS method (Left), and mass spectra of main species (Right)

Figure 3. Simultaneous identifications of charge variants and low molecular impurities in infliximab

Figure 4. Detection of low molecular weight impurities in the NISTmAb (upper panel), and the comparison of the results obtained from native SEC-MS (lower panel)

Figure 5. Charge variants analysis of intact (a) and IdeS digested (b) NISTmAb

Figure 6. Detection of partially unprocessed leader sequence in mAb 3

Table 1. Percentage of charge variant species under different stress conditions

	Acidic	Main	Basic
4ºC	15.70%	74.80%	9.50%
Low pH 1day	15.72%	74.64%	9.64%
Low pH 7day	13.85%	73.34%	12.81%
High pH 7day	80.92%	17.31%	1.77%
50ºC	21.35%	56.83%	21.82%
Light 1x	39.07%	41.30%	19.63

Figure 7. Comparison of charge variant separations under different stress conditions by SCX-MS (Left) and cIEF (Right)