Open AccessCrystallization and preliminary diffraction studies of prostaglandin E 2 ‐specific monoclonal antibody Fab fragment in the ligand complex
Author(s) -
Kurahashi Yuko,
Sugahara Mitsuaki,
Ago Hideo,
Aoyama Sae,
Takahashi Naoko,
Takio Koji,
Katsukawa Michiko,
Yamamoto Shozo,
Miyano Masashi
Publication year - 2008
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s174430910803131x
Subject(s) - prostaglandin , chemistry , prostaglandin e2 , monoclonal antibody , polyethylene glycol , crystallization , antibody , size exclusion chromatography , ligand (biochemistry) , molecule , biochemistry , biology , organic chemistry , receptor , immunology , enzyme , endocrinology
Prostaglandin E 2 is a major lipid mediator that regulates diverse biological processes. To elucidate how prostaglandin E 2 is recognized specifically by its antibody, the Fab fragment of a monoclonal anti‐prostaglandin E 2 antibody was prepared and its complex with prostaglandin E 2 was crystallized. The stable Fab–prostaglandin E 2 complex was prepared by gel‐filtration chromatography. Crystals were obtained by the microbatch method at 277 K using polyethylene glycol 4000 as a precipitant. A diffraction data set was collected to 2.2 Å resolution. The crystals belonged to space group P 2 1 2 1 2 1 , with unit‐cell parameters a = 70.3, b = 81.8, c = 82.2 Å. The asymmetric unit was suggested to contain one molecule of the Fab–prostaglandin E 2 complex, with a corresponding crystal volume per protein weight of 2.75 Å 3 Da −1 .