Table 2.
Dissociation binding constants and ΔG of binding for yeast surface displayed anti-fluorescein biotin binding scFv antibodies
| Mutant | No. of mutations, from 4-4-20 | Kd (± SD), pM | ΔGbind (± SD), kcal/mol | Δ ΔGbind from 4-4-20 |
| 4-4-20 | 0 | 1200 (± 300) | −12.16 (± 0.15) | 0 |
| 4M5.3 | 14 | 0.35 (± 0.08) | −17.0 (± 0.1) | −4.8 (± 0.2) |
| 4-4-20 F(L60)V | 1 | 540 (± 30) | −12.64 (± 0.03) | −0.5 (± 0.1) |
| 4-4-20 D(H31)H | 1 | 350 (± 40) | −12.89 (± 0.07) | −0.7 (± 0.1) |
| 4-4-20 I(H51)F | 1 | 2500 (± 300) | −11.73 (± 0.07) | 0.4 (± 0.1) |
| 4-4-20 S(H101)A | 1 | 720 (± 20) | −12.47 (± 0.02) | −0.3 (± 0.1) |
| 4-4-20 Y(H102)S | 1 | 320 (± 40) | −12.95 (± 0.07) | −0.8 (± 0.1) |
| 4-4-20 D(H106)E | 1 | 10,000 (± 5000) | −10.9 (± 0.3) | 1.3 (± 0.3) |
| 4-4-20 W(H108)L | 1 | 240 (± 40) | −13.1 (± 0.1) | −0.9 (± 0.1) |
| 4M5.3 V(L60)F | 13 | 0.7 (± 0.3) | −16.6 (± 0.3) | −4.4 (± 0.3) |
| 4M5.3 H(H31)D | 13 | 1.6 (± 0.2) | −16.08 (± 0.07) | −3.9 (± 0.1) |
| 4M5.3 F(H51)I | 13 | 2.0 (± 0.3) | −15.95 (± 0.09) | −3.8 (± 0.1) |
| 4M5.3 A(H101)S | 13 | 5.9 (± 0.4) | −15.31 (± 0.04) | −3.2 (± 0.1) |
| 4M5.3 S(H102)Y | 13 | 1.2 (± 0.1) | −16.27 (± 0.06) | −4.1 (± 0.1) |
| 4M5.3 E(H106)D | 13 | 0.7 (± 0.1) | −16.57 (± 0.08) | −4.4 (± 0.1) |
| 4M5.3 L(H108)W | 13 | 0.7 (± 0.1) | −16.6 (± 0.1) | −4.4 (± 0.1) |
| Min7 | 7 | 3.2 (± 0.3) | −15.67 (± 0.06) | −3.5 (± 0.1) |
| Min7 V(L60)F | 6 | N.D. | ||
| Min7 H(H31)D | 6 | 5.0 (± 0.4) | −15.41 (± 0.05) | −3.2 (± 0.1) |
| Min7 F(H51)I | 6 | 3.1 (± 0.8) | −15.7 (± 0.2) | −3.5 (± 0.2) |
| Min7 A(H101)S | 6 | N.D. | ||
| Min7 S(H102)Y | 6 | N.D. | ||
| Min7 E(H106)D | 6 | N.D. | ||
| Min7 L(H108)W | 6 | 2.3 (± 0.4) | −15.9 (0.1) | −3.7 (± 0.1) |
| CDR3 | 3 | 110 (± 10) | −13.57 (± 0.07) | −1.4 (± 0.1) |
| Non-CDR3 | 4 | 61 (± 7) | −13.93 (± 0.07) | −1.8 (± 0.1) |
| CDR3 + 108 | 4 | 21 (± 1) | −14.56 (± 0.03) | −2.4 (± 0.1) |
| Non-(CDR3 + 108) | 3 | 160 (± 20) | −13.36 (± 0.07) | −1.2 (± 0.1) |
| Contact | 3 | 38 (± 6) | −14.21 (± 0.09) | −2.0 (± 0.1) |
| Noncontact | 4 | 270 (± 60) | −13.1 (± 0.1) | −0.9 (± 0.2) |
| Early | 2 | 150 (± 20) | −13.38 (± 0.07) | −1.2 (± 0.1) |
| Late | 5 | 6.8 (± 1.4) | −15.2 (±.01) | −3.1 (± 0.2) |
N.D.: not determined. Error is indicated as standard deviation of replicate samples. The named mutants are defined as in Tables 1 and 3.