8CXC

Novel Anti-Mesothelin Antibodies Enable Crystallography of the Intact Mesothelin Ectodo- main and Engineering of Potent, T cell-engaging Bispecific Therapeutics

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	ALS BEAMLINE 5.0.2
Synchrotron site	ALS
Beamline	5.0.2
Temperature [K]	100
Detector technology	PIXEL
Collection date	2019-03-28
Detector	DECTRIS PILATUS 6M
Wavelength(s)	1.0
Spacegroup name	C 2 2 2
Unit cell lengths	185.303, 204.175, 123.114
Unit cell angles	90.00, 90.00, 90.00

Refinement procedure

Resolution	45.780 - 4.310
R-factor	0.2463
Rwork	0.243
R-free	0.29970
Structure solution method	MOLECULAR REPLACEMENT
Starting model (for MR)	4f33
RMSD bond length	0.011
RMSD bond angle	1.908
Data reduction software	HKL-2000
Data scaling software	SCALEPACK
Phasing software	PHASER
Refinement software	REFMAC (5.8.0267)

Data quality characteristics

	Overall	Inner shell	Outer shell
Low resolution limit [Å]	50.000	50.000	4.320
High resolution limit [Å]	4.250	11.490	4.250
Rmerge	0.121	0.068	0.784
Rmeas	0.151	0.089	0.979
Rpim	0.088	0.057	0.576
Total number of observations	34767
Number of reflections	13542	573	669
<I/σ(I)>	12.6
Completeness [%]	84.7	65.6	85.5
Redundancy	2.6	1.9	2.5
CC(1/2)		0.982	0.263

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, HANGING DROP	8	293	tris 8.0, zinc chloride, ammonium sulfate