1T3F

THREE DIMENSIONAL STRUCTURE OF A HUMANIZED ANTI-IFN-GAMMA FAB (HuZAF) IN P21 21 21 SPACE GROUP

Summary for 1T3F

• Entry DOI: 10.2210/pdb1t3f/pdb
• Related: 1B2W 1B4J 1T04
• Descriptor: Huzaf antibody light chain, Huzaf antibody heavy chain (3 entities in total)
• Functional Keywords: antibody engineering, humanized and chimeric antibody, fab, gamma-interferon, immune system
• Biological source: Homo sapiens (human); More
• Total number of polymer chains: 2
• Total formula weight: 46911.18

Authors

Bourne, P.C.,Terzyan, S.S.,Cloud, G.,Landolfi, N.F.,Vasquez, M.,Edmundson, A.B. (deposition date: 2004-04-26, release date: 2004-10-05, Last modification date: 2024-11-20)

Primary citation

Bourne, P.C.,Terzyan, S.S.,Cloud, G.,Landolfi, N.F.,Vasquez, M.,Edmundson, A.B.
Three-dimensional structures of a humanized anti-IFN-gamma Fab (HuZAF) in two crystal forms.
Acta Crystallogr.,Sect.D, 60:1761-1769, 2004

PubMed Abstract: Three-dimensional structures were determined for two crystal forms (orthorhombic P2(1)2(1)2(1) and monoclinic C2) of the Fab from the humanized version of a murine monoclonal antibody (AF2) that possesses binding and potent neutralizing activity against human interferon gamma (IFN-gamma). This humanized antibody (HuZAF; USAN name fontolizumab) is currently in phase II clinical trials for the treatment of Crohn's disease. HuZAF exhibits binding and IFN-gamma neutralizing capacities that closely approximate those of the original antibody. It is shown that HuZAF, whose VH domain was designed using a best-sequence-fit approach, is closer structurally to its mouse precursor than is a version whose VH was constructed using a human sequence with lower homology to the original mouse sequence. This work thus offers direct structural evidence in support of the best-sequence-fit approach and adds to previous results of biological and biochemical evaluations of distinctly engineered antibodies that also favored the use of a best-sequence-fit strategy. A second crystal type appeared during attempts to crystallize the Fab-IFN-gamma complex. The antibody-antigen complex that existed in solution dissociated in the crystallization mixture. A conformationally altered but unliganded HuZAF protein crystallized in a different space group (C2), with two Fab molecules in the asymmetric unit. In this crystal lattice, no space was available for accommodating the IFN-gamma antigen. Thus, there are currently three slightly different structures of the HuZAF Fab.

PubMed: 15388922
DOI: 10.1107/S0907444904018670

Experimental method

X-RAY DIFFRACTION (2 Å)