PDBsum entry 3dgg

Immune system

PDB id: 3dgg

Contents

Protein chains

217 a.a. *

212 a.a. *

Metals

_MG ×6

Waters ×367

* Residue conservation analysis

PDB id:

3dgg

Name:

Immune system

Title:

Crystal structure of fabox108

Structure:

Fabox108 light chain fragment. Chain: a, c. Synonym: vl. Engineered: yes. Fabox108 heavy chain fragment. Chain: b, d. Synonym: vh. Engineered: yes

Source:

Homo sapiens. Organism_taxid: 9606. Expressed in: homo sapiens. Expression_system_taxid: 9606. Expression_system_cell_line: hek293t. Expression_system_organ: human embryonic kidney cells.

Resolution:

2.30Å R-factor: 0.190 R-free: 0.246

Authors:

J.Ren,J.E.Nettleship,R.J.Owens,Oxford Protein Production Facility (Oppf)

Key ref:

J.E.Nettleship et al. (2008). A pipeline for the production of antibody fragments for structural studies using transient expression in HEK 293T cells. Protein Expr Purif, 62, 83-89. PubMed id: 18662785 DOI: 10.1016/j.pep.2008.06.017

Date:

13-Jun-08 Release date: 12-Aug-08

Protein chains

No UniProt id for this chain

Struc: 217 a.a.

Protein chains

No UniProt id for this chain

Struc: 212 a.a.

DOI no: 10.1016/j.pep.2008.06.017

Protein Expr Purif 62:83-89 (2008)

PubMed id: 18662785

A pipeline for the production of antibody fragments for structural studies using transient expression in HEK 293T cells.

J.E.Nettleship, J.Ren, N.Rahman, N.S.Berrow, D.Hatherley, A.N.Barclay, R.J.Owens.

ABSTRACT

We describe a pipeline for the rapid production of recombinant Fabs derived from mouse monoclonal antibodies suitable for use in structural studies. The pipeline is exemplified by the production of three Fabs derived from the monoclonal antibodies OX108 (anti-CD200 receptor), OX117 and OX119 (anti-SIRPgamma). Heavy and light chain variable domains were inserted into separate expression vectors containing resident constant regions using In-Fusion PCR cloning. Following transient co-expression in HEK 293T cells, secreted Fab fragments were purified by metal chelate chromatography and gel filtration using an automated procedure with yields of up to 4mg/L of cell culture. Following crystallization trials, diffracting crystals were obtained for the recombinant Fabs of OX108 and OX117, and their structures solved to 2.3A and 2.4A, respectively.