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. 2018 Jan 5;93(3):909–917. doi: 10.1002/jctb.5519

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© 2017 The Authors. Journal of Chemical Technology & Biotechnology published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

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JCTB-5519-FIG-0005-b — Peak height and peak wavelength against mAb clarified cell cultures. The dyes were spiked into wells containing clarified IgG1 mAb B from different time points. Supernatants were separated by centrifugation and filtration before measuring adding dye and measuring fluorescence. Comparison of decrease in wavelength at which highest fluorescence occurs (degree of blue shift) against increasing fluorescence intensity with increasing amount aggregates. Concentration of antibody in each well was 1 mg mL^‐1. (A) 50 µmol L^‐1 Bis‐ANS, ex/em‐ 390/450–600 nm, gain 70 (wavelength SD = 1 nm, peak intensity SD = 200 RFU); 5X SYPRO Orange, ex/em‐ 495/550–700 nm, gain 100 (wavelength SD = 1 nm, peak intensity SD = 207 RFU); (B) 3 µmol L^‐1 ProteoStat, ex/em‐ 530/560–700 nm (wavelength SD = 2 nm, peak intensity SD = 278 RFU); 50 µmol L^‐1 Thioflavin T, ex/em‐ 430/460–600 nm, gain 110 (wavelength SD = 1 nm, peak intensity SD = 122 RFU). Bis‐ANS SYPRO Orange Bis‐ANS SYPRO Orange ProteoStat Thioflavin ProteStat Thioflavin.

Inline graphic — Peak height and peak wavelength against mAb clarified cell cultures. The dyes were spiked into wells containing clarified IgG1 mAb B from different time points. Supernatants were separated by centrifugation and filtration before measuring adding dye and measuring fluorescence. Comparison of decrease in wavelength at which highest fluorescence occurs (degree of blue shift) against increasing fluorescence intensity with increasing amount aggregates. Concentration of antibody in each well was 1 mg mL^‐1. (A) 50 µmol L^‐1 Bis‐ANS, ex/em‐ 390/450–600 nm, gain 70 (wavelength SD = 1 nm, peak intensity SD = 200 RFU); 5X SYPRO Orange, ex/em‐ 495/550–700 nm, gain 100 (wavelength SD = 1 nm, peak intensity SD = 207 RFU); (B) 3 µmol L^‐1 ProteoStat, ex/em‐ 530/560–700 nm (wavelength SD = 2 nm, peak intensity SD = 278 RFU); 50 µmol L^‐1 Thioflavin T, ex/em‐ 430/460–600 nm, gain 110 (wavelength SD = 1 nm, peak intensity SD = 122 RFU). Bis‐ANS SYPRO Orange Bis‐ANS SYPRO Orange ProteoStat Thioflavin ProteStat Thioflavin.