6DN0

Retrofitted antibodies with stabilizing mutations: Herceptin scFv mutant with VH K30D and VL S52D.

Replaces:  4X4Y

Experimental 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)