9MI7

Crystal Structure of ADI-64597 ((human Fab, with substituted IgG1-CH1 (HC-L128R and K147R) and substituted kappa constant domain (LC-Q124E, V133Q, and T178R))

Experimental procedure

Experimental method	SINGLE WAVELENGTH
Source type	SYNCHROTRON
Source details	MAX IV BEAMLINE BioMAX
Synchrotron site	MAX IV
Beamline	BioMAX
Temperature [K]	100
Detector technology	PIXEL
Collection date	2021-05-12
Detector	DECTRIS EIGER X 16M
Wavelength(s)	0.9763
Spacegroup name	P 31
Unit cell lengths	65.983, 65.983, 97.431
Unit cell angles	90.00, 90.00, 120.00

Refinement procedure

Resolution	49.290 - 2.220
Rwork	0.180
R-free	0.23020
Structure solution method	MOLECULAR REPLACEMENT
Data reduction software	XDS
Data scaling software	Aimless
Phasing software	PHASER
Refinement software	PHENIX (1.19.2-4158)

Data quality characteristics

	Overall	Outer shell
Low resolution limit [Å]	49.290	2.300
High resolution limit [Å]	2.220	2.220
Number of reflections	23297	2261
<I/σ(I)>	11.6	1.4
Completeness [%]	99.7	97.6
Redundancy	10.4	10.5
CC(1/2)	0.998	0.542

Crystallization Conditions

crystal ID	method	pH	temperature	details
1	VAPOR DIFFUSION, SITTING DROP	7.5	295	ADI-64597 (human Fab) was received at concentration 16.5 mg/mL in a buffer containing 2 mM Tris-HCl pH 8.0 and 150 mM NaCl. PACT, BCS and JCSG+ screens (all from Molecular Dimensions Ltd.) were set up using a mosquito crystallisation robot (STP Labtech). Sitting drops of 150 nL protein and 150 nL reservoir solution were left to equilibrate against a 40 uL reservoir at 20 C. After a few days, needle-like crystals were obtained in several conditions. The crystal used for data collection was obtained in the BCS screen, condition B10 (0.1 M HEPES pH 7.5, 22% w/v PEG Smear Broad)