Sequential additive substitution mutagenesis of SakSTAR(K130T,K135R), SakSTAR(E80A,D82A,K130T,K135R), or SakSTAR(E65Q,K74Q,K130T,K135R) with selected other amino acids
| Variant . | Code . | Spec. Act. (kU/mg) . | Fibrinolytic potency in human plasma (C50 in μg/mL) . | Antibody absorption (%) . |
|---|---|---|---|---|
| SakSTAR | — | 130 | 0.18 | 95 |
| SakSTAR(K130T,K135R) | SY2 | 280 | 0.15 | 89 |
| SakSTAR(K74R,K130T,K135R) | SY4 | 310 | 0.17 | 76 |
| SakSTAR(K74Q,K130T,K135R) | SY41 | 190 | 0.18 | 62 |
| SakSTAR(E65A,K74Q,K130T,K135R) | SY48 | 170 | 0.20 | 55 |
| SakSTAR(E65Q,K74Q,K130T,K135R) | SY49 | 150 | — | 65 |
| SakSTAR(E65Q,K74Q,N95A,K130A,K135R) | SY72 | 220 | — | 58 |
| SakSTAR(K35A,E65Q,K74Q,K130A,K135R) | SY75 | 110 | 0.27 | 45 |
| SakSTAR(E80A,D82A,K130T,K135R) | SY6 | 250 | — | 80 |
| SakSTAR(K74R,E80A,D82A,K130T,K135R) | SY7 | 220 | 0.15 | 72 |
| SakSTAR(K74Q,E80A,D82A,K130T,K135R) | SY15 | 110 | 0.15 | 48 |
| SakSTAR(K35A,K74R,E80A,D82A,K130T,K135R) | SY17 | 160 | 0.19 | 68 |
| SakSTAR(K35A,K74Q,E80A,D82A,K130T,K135R) | SY28 | 120 | 0.18 | 48 |
| SakSTAR(E65D,K74R,E80A,D82A,K130T,K135R) | SY19 | 140 | 0.24 | 57 |
| SakSTAR(E65Q,K74R,E80A,D82A,K130T,K135R) | SY22 | 140 | 0.16 | 60 |
| SakSTAR(E65D,K74Q,E80A,D82A,K130T,K135R) | SY30 | 110 | 0.15 | 42 |
| SakSTAR(E65Q,K74Q,E80A,D82A,K130T,K135R) | SY47 | 120 | 0.16 | 42 |
| SakSTAR(K35A,E65D,K74R,E80A,D82A,K130T,K135R) | SY29 | 190 | 0.11 | 55 |
| SakSTAR(K35A,E65D,K74Q,E80A,D82A,K130T,K135R) | SY46 | 140 | 0.11 | 40 |
| SakSTAR(E65D,K74R,E80A,D82A,K130T,K135R,K136A) | SY35 | 100 | 0.19 | 45 |
| SakSTAR(K35A,E65D,K74R,E80A,D82A,K130T,K135R,K136A) | SY34 | 81 | — | 40 |
| SakSTAR(K35A,E65Q,K74Q,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R) | SY151 | 210 | 0.18 | 34 |
| SakSTAR(K35A,E65Q,K74R,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R) | SY155 | 180 | 0.21 | 36 |
| SakSTAR(E65Q,K74Q,K130T,K135R) | SY49 | 150 | 0.15 | 65 |
| SakSTAR(E65Q,K74Q,D82A,S84A,K130T,K135R) | SY50 | 170 | 0.19 | 45 |
| SakSTAR(E65Q,K74Q,T90A,E99D,T101S,K130A,K135R) | SY98 | 410 | 0.14 | 51 |
| SakSTAR(E65Q,K74Q,E108A,K109A,K130T,K135R) | SY83 | 180 | 0.18 | 50 |
| SakSTAR(E65Q,K74Q,D82A,S84A,E108A,K109A,K130T,K135R) | SY95 | 110 | 0.18 | 41 |
| SakSTAR(E65Q,K74Q,D82A,S84A,E108A,K109A,K130T,K135R,K136A,∇137K) | SY118 | 1500 | 0.15 | 30 |
| SakSTAR(E65Q,K74Q,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R,K136A,∇137K) | SY128 | 2900 | 0.23 | 28 |
| SakSTAR(K35A,E65Q,K74Q,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R,K136A,∇137K) | SY141 | 3700 | 0.19 | 24 |
| SakSTAR(K35A,E65Q,K74R,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R,K136A,∇137K) | SY145 | 5700 | 0.19 | 31 |
| Variant . | Code . | Spec. Act. (kU/mg) . | Fibrinolytic potency in human plasma (C50 in μg/mL) . | Antibody absorption (%) . |
|---|---|---|---|---|
| SakSTAR | — | 130 | 0.18 | 95 |
| SakSTAR(K130T,K135R) | SY2 | 280 | 0.15 | 89 |
| SakSTAR(K74R,K130T,K135R) | SY4 | 310 | 0.17 | 76 |
| SakSTAR(K74Q,K130T,K135R) | SY41 | 190 | 0.18 | 62 |
| SakSTAR(E65A,K74Q,K130T,K135R) | SY48 | 170 | 0.20 | 55 |
| SakSTAR(E65Q,K74Q,K130T,K135R) | SY49 | 150 | — | 65 |
| SakSTAR(E65Q,K74Q,N95A,K130A,K135R) | SY72 | 220 | — | 58 |
| SakSTAR(K35A,E65Q,K74Q,K130A,K135R) | SY75 | 110 | 0.27 | 45 |
| SakSTAR(E80A,D82A,K130T,K135R) | SY6 | 250 | — | 80 |
| SakSTAR(K74R,E80A,D82A,K130T,K135R) | SY7 | 220 | 0.15 | 72 |
| SakSTAR(K74Q,E80A,D82A,K130T,K135R) | SY15 | 110 | 0.15 | 48 |
| SakSTAR(K35A,K74R,E80A,D82A,K130T,K135R) | SY17 | 160 | 0.19 | 68 |
| SakSTAR(K35A,K74Q,E80A,D82A,K130T,K135R) | SY28 | 120 | 0.18 | 48 |
| SakSTAR(E65D,K74R,E80A,D82A,K130T,K135R) | SY19 | 140 | 0.24 | 57 |
| SakSTAR(E65Q,K74R,E80A,D82A,K130T,K135R) | SY22 | 140 | 0.16 | 60 |
| SakSTAR(E65D,K74Q,E80A,D82A,K130T,K135R) | SY30 | 110 | 0.15 | 42 |
| SakSTAR(E65Q,K74Q,E80A,D82A,K130T,K135R) | SY47 | 120 | 0.16 | 42 |
| SakSTAR(K35A,E65D,K74R,E80A,D82A,K130T,K135R) | SY29 | 190 | 0.11 | 55 |
| SakSTAR(K35A,E65D,K74Q,E80A,D82A,K130T,K135R) | SY46 | 140 | 0.11 | 40 |
| SakSTAR(E65D,K74R,E80A,D82A,K130T,K135R,K136A) | SY35 | 100 | 0.19 | 45 |
| SakSTAR(K35A,E65D,K74R,E80A,D82A,K130T,K135R,K136A) | SY34 | 81 | — | 40 |
| SakSTAR(K35A,E65Q,K74Q,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R) | SY151 | 210 | 0.18 | 34 |
| SakSTAR(K35A,E65Q,K74R,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R) | SY155 | 180 | 0.21 | 36 |
| SakSTAR(E65Q,K74Q,K130T,K135R) | SY49 | 150 | 0.15 | 65 |
| SakSTAR(E65Q,K74Q,D82A,S84A,K130T,K135R) | SY50 | 170 | 0.19 | 45 |
| SakSTAR(E65Q,K74Q,T90A,E99D,T101S,K130A,K135R) | SY98 | 410 | 0.14 | 51 |
| SakSTAR(E65Q,K74Q,E108A,K109A,K130T,K135R) | SY83 | 180 | 0.18 | 50 |
| SakSTAR(E65Q,K74Q,D82A,S84A,E108A,K109A,K130T,K135R) | SY95 | 110 | 0.18 | 41 |
| SakSTAR(E65Q,K74Q,D82A,S84A,E108A,K109A,K130T,K135R,K136A,∇137K) | SY118 | 1500 | 0.15 | 30 |
| SakSTAR(E65Q,K74Q,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R,K136A,∇137K) | SY128 | 2900 | 0.23 | 28 |
| SakSTAR(K35A,E65Q,K74Q,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R,K136A,∇137K) | SY141 | 3700 | 0.19 | 24 |
| SakSTAR(K35A,E65Q,K74R,D82A,S84A,T90A,E99D,T101S,E108A,K109A,K130T,K135R,K136A,∇137K) | SY145 | 5700 | 0.19 | 31 |
Specific activity (Spec. Act.) ≥ 100 kU/mg is represented in bold type. Absorption of antibodies (in %) from pooled immunized patient plasma; values ≤ 60% are represented in bold type. C50: concentration (in μg/mL) causing 50% clot lysis in 2 hours.