4KPH

Structure of the Fab fragment of N62, a protective monoclonal antibody to the nonreducing end of Francisella tularensis O-antigen

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	ROTATING ANODE
Source details	RIGAKU RU300
Temperature [K]	100
Detector technology	IMAGE PLATE
Collection date	2012-05-18
Detector	RIGAKU RAXIS IV
Wavelength(s)	1.5418
Spacegroup name	P 21 21 21
Unit cell lengths	68.874, 87.076, 153.589
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	14.870 - 2.590
R-factor	0.225
Rwork	0.222
R-free	0.26800
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	3OZ9 (light chain) and pdb entry 3D9A (heavy chain)
RMSD bond length	0.002
RMSD bond angle	0.666
Data reduction software	DENZO
Data scaling software	SCALEPACK
Phasing software	PHASER (2.5.0)
Refinement software	PHENIX (1.8_1069)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	15.000	15.000	2.690
High resolution limit [Å]	2.593	5.520	2.600
Rmerge	0.068	0.033	0.325
Number of reflections	28828
<I/σ(I)>	14.6
Completeness [%]	98.8	97.6	98.1
Redundancy	3.8	3.7	3.7

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	6.5	290	Microseeding by mixing 0.5 ul of Fab (12 mg/ml in 20 mM Tris-HCl pH 7.5, 150 mM NaCl, 0.02% NaN3) with 0.5 ul of reservoir solution [0.1 M bistris (pH 6.5), 0.2 M ammonium acetate, 25% w/v PEG 3350] and streaking after equilibration, VAPOR DIFFUSION, HANGING DROP, temperature 290K