Table 2.
Comparison of Amenability Outcomes Between a GLP-HEK Assay and In-House Assays
| Assay | Benjamin et al.10 | Lukas et al.39 | Oommen et al.36 | Lenders et al.26 | Benjamin et al.10 | Lukas et al.39 | Oommen et al.36 | Lenders et al.26 | ||||
|---|---|---|---|---|---|---|---|---|---|---|---|---|
| Amino Acid | w/o DGJ | Absolute Increase %WT, (n-Fold) | w/o DGJ | Absolute Increase %WT, (n-Fold) | w/o DGJ | Absolute Increase %WT, (n-Fold) | w/o DGJ | Absolute Increase %WT, (n-Fold) | Amenable (Yes/No) | |||
| p.L3P | 71.9 | 20.3 (1.3) | 117.7 | 11.3 (1.1) | 69.4 | 37.1 (1.5) | yes | yes | yes | |||
| p.A13T | 51.7 | 10.4 (1.2) | 61.9 | 19.6 (1.3) | yes | yes | ||||||
| p.A15G | 19.0 | 9.0 (1.5) | 15.5 | 15.5 (2.0) | yes | yes | ||||||
| p.A15T | 39.1 | 18.4 (1.5) | 12.4 | 85.2 (7.8) | yes | yes | ||||||
| p.A20D | 4.3 | 5.7 (2.4) | 5.8 | 10.6 (2.8) | yes | yes | ||||||
| p.A20P | 11.5 | 4.4 (1.4) | 13.6 | 22.6 (2.7) | yes | yes | ||||||
| p.W24R | 52.6 | 10.9 (1.2) | 50.5 | 20.6 81.4) | yes | yes | ||||||
| p.D33G | 29.3 | 41.3 (2.4) | 37.4 | 24.6 (1.7) | yes | yes | ||||||
| p.N34S | 0.6 | 16.1 (29.2) | 2.0 | 14.1 (8.2) | 0.9 | 1.5 (2.7) | yes | yes | no | |||
| p.L36W | 0.7 | 15.9 (23.7) | 2.3 | 20 (9.7) | yes | yes | ||||||
| p.A37T | 48.9 | 47.5 (2.0) | 69.6 | 63.3 (1.9) | yes | yes | ||||||
| p.M42L | 38.8 | 21.3 (1.5) | 16.5 | 26.5 (2.6) | yes | yes | ||||||
| p.M42V | 0.5 | 3.8 (8.6) | 0 | 7.2 (n/c) | yes | yes | ||||||
| p.M42T | 2.5 | 17.8 (8.1) | 2.9 | 18.5 (7.4) | yes | yes | ||||||
| p.H46P | 31 | 75.9 (3.4) | 40.1 | 58.7 (2.5) | yes | yes | ||||||
| p.M51K | 6.3 | 15.8 (3.5) | 0 | 8.7 (n/c) | yes | yes | ||||||
| p.M51I | 22.3 | 24.8 82.1) | 37.4 | 24.6 (1.7) | yes | yes | ||||||
| p.C56Y | N/A | 7.3 (n/c) | 0.0 | 6.6 (n/c) | yes | yes | ||||||
| p.Q57L | 71.6 | 21.7 (1.3) | 52.9 | 28.4 (1.5) | yes | yes | ||||||
| p.E59K | 8.6 | 8.9 (2.0) | 2.2 | 16.3 (8.4) | yes | yes | ||||||
| p.P60L | 21.7 | 39.3 (2.8) | 15.6 | 17.5 (2.1) | yes | yes | ||||||
| p.E66G | 34.2 | 11.9 (1.4) | 28.9 | 37.0 (2.3) | yes | yes | ||||||
| p.E66K | 4.8 | 8.1 (2.7) | 6.8 | 11.5 (2.7) | 6.9 | 8.8 (2.3) | yes | yes | yes | |||
| p.M72I | 54.7 | 22.8 (1.4) | 106.5 | 101.9 (2.0) | yes | yes | ||||||
| p.A73V | 53.6 | 33.3 (1.6) | 44 | 20.7 (1.5) | yes | yes | ||||||
| p.D83N | 69.2 | 23.8 (1.3) | 62.9 | 8.7 (1.1) | 187.5 | 54.3 (1.3) | yes | yes | yes | |||
| p.G85M | 7.9 | 4.0 (1.5) | 6.5 | 15.1 (3.3) | yes | yes | ||||||
| p.G85S | 12.4 | 6.7 (1.5) | 7.6 | 12.2 (2.6) | yes | yes | ||||||
| p.I91T | 0.9 | 11.7 (14.0) | 0.7 | 6.3 (10.0) | yes | yes | ||||||
| p.S102L | 19.9 | 42.9 (3.2) | 71.6 | 7.3 (1.1) | yes | yes | ||||||
| p.A108T | 57.1 | 23.7 (1.4) | 73.3 | 11.2 (1.15) | yes | no | ||||||
| p.R112H | 2.6 | 14.8 (6.7) | 1.6 | 17.8 (12.1) | yes | yes | ||||||
| p.R118C | 24.0 | 5.5 (1.3) | 5.3 | 5.8 (2.1) | 45.4 | 20.1 (1.4) | yes | yes | yes | |||
| p.A121T | 18.9 | 49.0 (3.6) | 50 | 5.5 (1.1) | yes | yes | ||||||
| p.S126G | 83.7 | 30.2 (1.4) | 51.3 | 16.1 (1.3) | 143.7 | 26.9 (1.19) | yes | yes | no | |||
| p.G128E | 45.2 | 13.4 (1.3) | 26.4 | 17.4 (1.7) | yes | yes | ||||||
| p.A135V | 0 | 3.7 (n/c) | 0 | 6.9 (n/c) | 0.1 | 0.1 (2.0) | yes | yes | no | |||
| p.D136E | 1.4 | 11.5 (9.2) | 0 | 31.3 (n/c) | yes | yes | ||||||
| p.N139S | 65.5 | 13.6 (1.2) | 64.6 | 9.7 (1.2) | 70.1 | 47.6 (1.7) | 103.4 | 24.0 (1.2) | yes | yes | yes | yes |
| p.A143T | 21.4 | 22.4 (2.0) | 31.3 | 18.1 (1.6) | yes | yes | ||||||
| p.G144D | 50.2 | 26.3 (1.5) | 28.8 | 16.1 (1.6) | yes | yes | ||||||
| p.P146S | 41.9 | 22.2 (1.5) | 17.9 | 22.7 (2.3) | yes | yes | ||||||
| p.A156V | 1.2 | 11.6 (10.7) | 4.3 | 12.5 (3.9) | yes | yes | ||||||
| p.W162G | 0.8 | 5.1 (7.4) | 0 | 5.2 (n/c) | 1.4 | 1.5 (2.1) | yes | yes | no | |||
| p.D165H | 1.3 | 7.0 (6.4) | 3.4 | 8.5 (3.5) | yes | yes | ||||||
| p.D175E | 44.3 | 9.1 (1.2) | 57.9 | 9.6 (1.17) | yes | no | ||||||
| p.G183A | 22.4 | 34.0 (2.5) | 10 | 36.6 (4.6) | yes | yes | ||||||
| p.M187V | 1.3 | 13.6 (11.5) | 22.8 | 44.2 (2.9) | 12.2 | 26.5 (3.2) | yes | yes | yes | |||
| p.M187I | 5.1 | 25.6 (6.0) | 3.1 | 28.1 (10.1) | yes | yes | ||||||
| p.I198T | 64.7 | 30.8 (1.5) | 38.7 | 11.7 (1.3) | 58.3 | 58.5 (2.0) | yes | yes | yes | |||
| p.E203V | 43.0 | 21.5 (1.5) | 144.6 | 72.1 (1.5) | yes | yes | ||||||
| p.P205T | 14.4 | 34.4 (3.4) | 10.2 | 60.2 (6.9) | yes | yes | ||||||
| p.P210S | 75.2 | 38.2 (1.5) | 41.9 | 49.5 (2.2) | yes | yes | ||||||
| p.K213M | 43.2 | 12.7 (1.3) | 31.5 | 12.9 (1.4) | 61.6 | 21.6 (1.4) | yes | yes | yes | |||
| p.P214S | 22.4 | 60.1 (3.7) | 18.1 | 43.5 (3.4) | yes | yes | ||||||
| p.P214L | 33 | 58.6 (2.8) | 19.4 | 44.7 (3.3) | yes | yes | ||||||
| p.N215D | 43.8 | 14.5 (1.3) | 41.1 | 22.4 (1.5) | yes | yes | ||||||
| p.N215S | 15.6 | 20.0 (2.3) | 36.7 | 24.9 (1.7) | 27.1 | 28.1 (2.0) | 26.8 | 57.5 (3.1) | yes | yes | yes | yes |
| p.Y216C | 2 | 18.7 (10.4) | 2.3 | 23.6 (11.2) | yes | yes | ||||||
| p.I219T | 55.8 | 37.8 (1.7) | 53.3 | 32 (1.6) | yes | yes | ||||||
| p.R220Q | 45.2 | 16.5 (1.4) | 115.5 | 38.7 (1.3) | yes | yes | ||||||
| p.N224S | 10.3 | 19.4 (2.9) | 31.1 | 51.1 (2.6) | yes | yes | ||||||
| p.H225D | 43.8 | 66.8 (2.5) | 32.2 | 28.3 (1.9) | yes | yes | ||||||
| p.N228S | 124.5 | 44.7 (1.4) | 59.5 | 11.1 (1.2) | 89.6 | 54.9 (1.6) | yes | yes | yes | |||
| p.I232T | 15 | 70.0 (5.7) | 11.5 | 50.1 (5.4) | yes | yes | ||||||
| p.S238N | 37.1 | 59.3 (2.6) | 36 | 58.3 (2.6) | yes | yes | ||||||
| p.I239T | 37.7 | 55.1 (2.5) | 26.6 | 58.3 (3.2) | yes | yes | ||||||
| p.K240N | N/A | > 3% (≥1.2) | 33.4 | 24.4 (1.7) | yes | yes | ||||||
| p.I242N | 7.6 | 59.8 (59.8) | 3.1 | 46.7 (16.1) | yes | yes | ||||||
| p.L243F | 7.9 | 34.4 (5.4) | 11.4 | 59.4 (6.2) | yes | yes | ||||||
| p.W245G | 44.1 | 18.9 (1.4) | 51.3 | 229.9 (5.5) | yes | yes | ||||||
| p.N249K | 17.9 | 17.3 (2.0) | 23.7 | 30.9 (2.3) | yes | yes | ||||||
| p.Q250P | 24.8 | 33.9 (2.4) | 18.5 | 42.5 (3.3) | yes | yes | ||||||
| p.I253T | 38.9 | 41.3 (2.1) | 73 | 42.8 (1.6) | yes | yes | ||||||
| p.I253S | 3.3 | 27.9 (9.5) | 4.4 | 49 (12.1) | yes | yes | ||||||
| p.P259R | 23.3 | 37 (2.6) | 20.5 | 19.5 (2.0) | yes | yes | ||||||
| p.N263S | 15.8 | 64.7 (5.1) | 6.7 | 57.8 (9.7) | yes | yes | ||||||
| p.D264Y | 0.5 | 5.7 (12.9) | 0 | 5.4 (n/c) | 2.8 | 8.6 (4.0) | yes | yes | yes | |||
| p.V269M | 4.4 | 21.5 (5.9) | 0 | 17.3 (n/c) | yes | yes | ||||||
| p.V269A | 0 | 7.8 (n/c) | 9 | 36 (5.0) | yes | yes | ||||||
| p.G271D | 1.5 | 30.7 (21.5) | 3.1 | 34.8 (12.3) | yes | yes | ||||||
| p.S276N | 2.3 | 7.0 (3.9) | 4.6 | 40.4 (9.9) | yes | yes | ||||||
| p.T282I | 5.2 | 18.5 (4.6) | 5 | 42.7 (9.5) | 12.6 | 9.2 (1.7) | yes | yes | yes | |||
| p.M284V | 25.2 | 37.9 (2.5) | 16.2 | 27.2 (2.7) | yes | yes | ||||||
| p.M290L | 58.6 | 52.8 (1.9) | 69.4 | 53.1 (1.8) | yes | yes | ||||||
| p.A291T | 16.5 | 24.0 (2.5) | 13.2 | 42.5 (4.2) | yes | yes | ||||||
| p.L294S | 0 | 4.9 (n/c) | 0 | 12.4 (n/c) | 0.2 | 0.3 (2.5) | yes | yes | no | |||
| p.R301G | 19.1 | 45.6 (3.4) | 19.3 | 37.2 (2.9) | yes | yes | ||||||
| p.R301Q | 5.5 | 39.0 (8.1) | 8.5 | 39.5 (5.6) | yes | yes | ||||||
| p.R301P | 0 | 4.2 (n/c) | 0 | 5 (n/c) | yes | yes | ||||||
| p.S304N | 94.1 | 27.7 (1.3) | 77.8 | 12.5 (1.16) | yes | no | ||||||
| p.S304T | 76.4 | 40.5 (1.5) | 56.7 | 23.4 (1.4) | yes | yes | ||||||
| p.L310F | 0.8 | 10.8 (14.5) | 1.6 | 26.1 (17.3) | 1.8 | 24.5 (14.6) | yes | yes | yes | |||
| p.L311V | 2 | 16.0 (9.0) | 1.9 | 38.2 (21.1) | yes | yes | ||||||
| p.D313G | 25.5 | 14.9 (1.6) | 25.1 | 25.4 (2.0) | yes | yes | ||||||
| p.D313Y | 59 | 21.9 (1.4) | 83.9 | 16.4 (1.2) | 54.2 | 28.9 (1.5) | yes | yes | yes | |||
| p.V316I | 92.1 | 34.0 (1.4) | 99.8 | 33.4 (1.3) | yes | yes | ||||||
| p.I317T | 6.5 | 17.0 (3.4) | 2.7 | 15.2 (6.7) | yes | yes | ||||||
| p.I319T | 10.3 | 17.7 (2.7) | 20.2 | 38.1 (2.9) | yes | yes | ||||||
| p.N320I | 1.4 | 15.7 (12.2) | 2 | 29.8 (15.9) | 6.0 | 14.3 (3.4) | yes | yes | yes | |||
| p.Q321H | 1.9 | 17.9 (10.4) | 3.3 | 22.0 (7.7) | yes | yes | ||||||
| p.D322N | 36.1 | 10.3 (1.3) | 17.8 | 13.3 (1.8) | yes | yes | ||||||
| p.G325S | 24.7 | 37.8 (2.5) | 25.6 | 29.8 (2.2) | yes | yes | ||||||
| p.Q327E | 22.9 | 27.5 (2.2) | 21.5 | 59.4 (3.8) | yes | yes | ||||||
| p.G328A | 6.9 | 21.8 (4.2) | 6.2 | 23.8 (4.8) | yes | yes | ||||||
| p.G334E | 86.7 | 18.8 81.2) | 57.5 | 23.4 (1.4) | yes | yes | ||||||
| p.P343L | 36.6 | 13.0 (1.4) | 39.6 | 22.6 (1.6) | yes | yes | ||||||
| p.S345P | 0 | 3.7 (n/c) | 0 | 9.7 (n/c) | yes | yes | ||||||
| p.R356W | 11 | 38.1 (4.5) | 16.9 | 45.8 (3.7) | yes | yes | ||||||
| p.R356Q | 36.1 | 39.0 (2.1) | 33.3 | 33.0 (2.0) | yes | yes | ||||||
| p.G360C | 8.7 | 8.1 (1.9) | 16.2 | 16.7 (2.0) | 5.3 | 9.9 (2.9) | yes | yes | yes | |||
| p.R363H | 20 | 30.5 (2.5) | 31.9 | 26.0 (1.8) | yes | yes | ||||||
| p.A368T | 54.6 | 18.0 (1.3) | 79.6 | 21.3 (1.3) | yes | yes | ||||||
| p.T385A | 57.3 | 16.1 (1.3) | 28.6 | 17.3 (1.6) | 68.1 | 9.6 (1.1) | yes | yes | no | |||
| p.G395A | 24.4 | 6.3 (1.3) | 13.4 | 5.2 (1.4) | yes | yes | ||||||
| p.L403S | 14.0 | 4.9 (1.4) | 13.2 | 13.0 (2.0) | yes | yes | ||||||
| p.T410I | 0.4 | 11.8 (30.5) | 2.3 | 13.8 (7.0) | yes | yes | ||||||
| p.E418G | 67.5 | 21.9 (1.3) | 74.6 | 14.5 (1.2) | 140.7 | 51.8 (1.4) | yes | yes | yes | |||
| p.M421V | 83.0 | 25.6 81.3) | 58.3 | 26.0 (1.5) | yes | yes | ||||||
Only amenable mutations according to the GLP-HEK assay were included. According to the GLP-HEK assay, an amenable mutation responds with an absolute increase by ≥3% of wild-type AGAL activity and a ≥1.2-fold increase of baseline enzymatic activity in the presence of 10 μM migalastat. w/o, without; WT, wild-type; NA, not available; n/c, not calculated.