6DN0
Retrofitted antibodies with stabilizing mutations: Herceptin scFv mutant with VH K30D and VL S52D.
Replaces: 4X4YExperimental procedure
Experimental method | SINGLE WAVELENGTH |
Source type | SYNCHROTRON |
Source details | AUSTRALIAN SYNCHROTRON BEAMLINE MX2 |
Synchrotron site | Australian Synchrotron |
Beamline | MX2 |
Temperature [K] | 100 |
Detector technology | CCD |
Collection date | 2016-08-13 |
Detector | ADSC QUANTUM 315r |
Wavelength(s) | 0.9537 |
Spacegroup name | P 31 2 1 |
Unit cell lengths | 91.981, 91.981, 114.677 |
Unit cell angles | 90.00, 90.00, 120.00 |
Refinement procedure
Resolution | 39.830 - 2.000 |
R-factor | 0.2063 |
Rwork | 0.205 |
R-free | 0.23380 |
Structure solution method | MOLECULAR REPLACEMENT |
Starting model (for MR) | 4x4x |
RMSD bond length | 0.010 |
RMSD bond angle | 1.322 |
Data reduction software | MOSFLM |
Data scaling software | Aimless (0.5.17) |
Phasing software | PHASER (2.5.7) |
Refinement software | REFMAC (5.8.0222) |
Data quality characteristics
Overall | Outer shell | |
Low resolution limit [Å] | 39.830 | 2.050 |
High resolution limit [Å] | 2.000 | 2.000 |
Rmerge | 0.101 | 1.018 |
Rmeas | 0.106 | 1.064 |
Rpim | 0.031 | 0.304 |
Total number of observations | 458269 | |
Number of reflections | 38572 | 2773 |
<I/σ(I)> | 14.6 | |
Completeness [%] | 99.9 | 98.8 |
Redundancy | 11.9 | 11.7 |
CC(1/2) | 0.999 | 0.855 |
Crystallization Conditions
crystal ID | method | pH | temperature | details |
1 | VAPOR DIFFUSION, SITTING DROP | 4.6 | 293 | Equal volumes of protein (8.0 mg/mL in 25 mM Tris (pH 8.0)) were combined with an equal volume of well solution (3.5 M sodium formate, 100 mM sodium acetate (pH 4.6) |