From: Determination of immunogenic proteins in biopharmaceuticals by UHPLC–MS amino acid analysis
Within-run, n = 6 | Between-run, n = 12 | |||||
---|---|---|---|---|---|---|
Mean found concentration (µM) | RE (% Nom.) | RSD (%) | Mean found concentration (µM) | RE (% Nom.) | RSD (%) | |
Glycine | ||||||
5 µM | 4.50 | − 9.00 | 3.14 | 4.90 | − 2.00 | 9.72 |
50 µM | 50.45 | 0.90 | 7.99 | 49.07 | − 1.86 | 5.59 |
250 µM | 254.55 | 1.82 | 6.81 | 240.60 | − 3.76 | 6.69 |
Alanine | ||||||
5 µM | 4.70 | − 6.00 | 3.01 | 4.55 | − 9.00 | 8.98 |
50 µM | 49.25 | − 1.50 | 0.43 | 48.87 | − 2.26 | 1.70 |
250 µM | 256.30 | 2.52 | 2.59 | 247.52 | − 0.99 | 3.05 |
Serine | ||||||
5 µM | 4.55 | − 9.00 | 7.77 | 4.68 | − 6.40 | 6.39 |
50 µM | 49.15 | − 1.70 | 2.45 | 48.45 | − 3.10 | 4.04 |
250 µM | 253.50 | 1.40 | 2.79 | 253.1 | 1.24 | 2.77 |
Proline | ||||||
5 µM | 4.80 | − 4.00 | 2.95 | 4.78 | − 4.40 | 2.78 |
50 µM | 49.90 | − 0.20 | 7.09 | 50.63 | 1.26 | 3.85 |
250 µM | 259.75 | 3.90 | 2.37 | 252.35 | 0.94 | 3.77 |
Valine | ||||||
5 µM | 4.78 | − 4.4 | 5.51 | 4.73 | − 5.40 | 4.56 |
50 µM | 48.90 | − 2.20 | 2.60 | 50.65 | 1.30 | 4.12 |
250 µM | 249.85 | − 0.06 | 0.25 | 245.90 | − 1.64 | 2.00 |
Threonine | ||||||
5 µM | 4.95 | − 1.00 | 7.14 | 4.62 | − 7.60 | 7.43 |
50 µM | 49.95 | − 0.10 | 0.71 | 49.05 | − 1.90 | 3.23 |
250 µM | 251.25 | 0.50 | 1.21 | 240.7 | − 3.72 | 3.77 |
Cysteine | ||||||
2.5 µM | 2.60 | 4.00 | 5.44 | 2.53 | 1.20 | 4.11 |
25 µM | 25.05 | 0.20 | 3.11 | 24.73 | − 1.08 | 4.22 |
125 µM | 122.78 | − 1.78 | 2.85 | 120.44 | − 3.65 | 2.45 |
Isoleucine | ||||||
5 µM | 5.05 | 1.00 | 1.40 | 4.98 | − 0.40 | 3.90 |
50 µM | 49.60 | − 0.80 | 3.42 | 50.45 | 0.90 | 2.73 |
250 µM | 258.35 | 3.34 | 4.79 | 247.37 | − 1.05 | 4.24 |
Leucine | ||||||
5 µM | 4.85 | − 3.00 | 1.46 | 4.72 | − 5.60 | 4.91 |
50 µM | 48.95 | − 2.10 | 3.61 | 48.92 | − 2.16 | 1.67 |
250 µM | 252.95 | 1.18 | 5.40 | 243.43 | − 2.63 | 4.32 |
Asparagine | ||||||
5 µM | 5.30 | 6.00 | 8.22 | 4.98 | − 0.40 | 11.57 |
50 µM | 53.75 | 7.50 | 0.13 | 52.60 | 5.20 | 1.93 |
250 µM | 249.75 | − 0.10 | 2.58 | 242.53 | − 2.99 | 2.64 |
Aspartic acid | ||||||
5 µM | 4.65 | − 7.00 | 1.52 | 4.43 | − 11.40 | 11.31 |
50 µM | 51.35 | 2.70 | 4.82 | 47.82 | − 4.36 | 6.73 |
250 µM | 262.40 | 4.96 | 11.97 | 247.03 | − 1.19 | 7.91 |
Glutamine | ||||||
5 µM | 4.85 | − 3.00 | 1.46 | 4.53 | − 9.40 | 5.52 |
50 µM | 53.25 | 6.50 | 2.26 | 52.48 | 4.96 | 2.20 |
250 µM | 245.15 | − 1.94 | 5.51 | 245.32 | − 1.87 | 3.18 |
Lysine | ||||||
5 µM | 4.95 | − 1.00 | 6.70 | 4.85 | − 3.00 | 6.22 |
50 µM | 52.00 | 4.00 | 1.90 | 51.62 | 3.24 | 3.29 |
250 µM | 252.70 | 1.08 | 0.39 | 246.60 | − 1.36 | 3.71 |
Glutamic acid | ||||||
5 µM | 4.75 | − 5.00 | 1.49 | 4.67 | − 6.60 | 1.75 |
50 µM | 50.20 | 0.40 | 8.48 | 48.28 | − 3.44 | 4.07 |
250 µM | 261.10 | 4.44 | 6.07 | 260.27 | 4.11 | 3.15 |
Methionine | ||||||
5 µM | 4.75 | − 5.00 | 1.49 | 4.97 | − 0.60 | 4.35 |
50 µM | 50.20 | 0.40 | 5.92 | 49.03 | − 1.94 | 4.25 |
250 µM | 253.20 | 1.28 | 6.26 | 245.90 | − 1.64 | 3.88 |
Histidine | ||||||
5 µM | 4.63 | − 7.40 | 10.65 | 4.72 | − 5.60 | 9.40 |
50 µM | 50.60 | 1.12 | 2.80 | 49.05 | − 1.90 | 4.71 |
250 µM | 251.70 | 0.68 | 2.59 | 249.07 | − 0.37 | 2.22 |
Phenylalanine | ||||||
5 µM | 4.85 | − 3.00 | 1.46 | 4.80 | − 4.00 | 1.86 |
50 µM | 47.85 | − 4.30 | 4.29 | 46.45 | − 7.10 | 3.56 |
250 µM | 258.00 | 3.20 | 1.21 | 249.48 | − 0.21 | 3.79 |
Arginine | ||||||
5 µM | 4.95 | − 1.00 | 4.29 | 4.62 | − 7.60 | 7.17 |
50 µM | 48.45 | − 3.10 | 2.19 | 48.77 | − 2.46 | 6.86 |
250 µM | 258.15 | 3.26 | 0.36 | 245.08 | − 1.97 | 5.06 |
Tyrosine | ||||||
5 µM | 4.77 | − 4.60 | 1.21 | 4.63 | − 7.40 | 3.53 |
50 µM | 49.30 | − 1.40 | 3.44 | 47.22 | − 5.56 | 4.11 |
250 µM | 256.05 | 2.42 | 6.43 | 246.23 | − 1.51 | 5.26 |
Tryptophan | ||||||
5 µM | 5.15 | 3.00 | 1.37 | 4.98 | − 0.40 | 6.14 |
50 µM | 49.55 | − 0.90 | 2.14 | 50.40 | 0.80 | 2.70 |
250 µM | 263.90 | 5.56 | 9.59 | 254.52 | 1.81 | 5.57 |