The structure of a GFP-based antibody (fluorobody) to TLH, a toxin from Vibrio parahaemolyticus

• Published in: Acta crystallographica. Section F, Structural biology communications Vol. 71; no. 7; pp. 913 - 918
• Main Authors: Chen, Yaoguang, Huang, Xiaocheng, Wang, Rongzhi, Wang, Shihua, Shi, Ning
• Format: Journal Article
• Language: English
• Published: 5 Abbey Square, Chester, Cheshire CH1 2HU, England International Union of Crystallography 01.07.2015; Wiley Subscription Services, Inc
• Subjects: Amino Acid Sequence; Antibodies, Bacterial - chemistry; Antibodies, Bacterial - metabolism; Bacterial Proteins - chemistry; Bacterial Proteins - isolation & purification; Bacterial Proteins - metabolism; CDR3; fluorobody; GFP scaffold; Green Fluorescent Proteins - chemistry; Green Fluorescent Proteins - metabolism; Hemolysin Proteins - chemistry; Hemolysin Proteins - isolation & purification; Hemolysin Proteins - metabolism; Molecular Sequence Data; Protein Structure, Secondary; Research Communications; TLH; Vibrio parahaemolyticus; TLH; CDR3; fluorobody; GFP scaffold
• ISSN: 2053-230X; 2053-230X
• DOI: 10.1107/S2053230X15008845

A fluorobody is a manmade hybrid molecule that is composed of green fluorescent protein (GFP) and a fragment of antibody, which combines the affinity and specificity of an antibody with the visibility of a GFP. It is able to provide a real‐time indication of binding while avoiding the use of tags and secondary binding reagents. Here, the expression, purification and crystal structure of a recombinant fluorobody for TLH (thermolabile haemolysin), a toxin from the lethal food‐borne disease bacterium Vibrio parahaemolyticus, are presented. This is the first structure of a fluorobody to be reported. Crystals belonging to space group P43212, with unit‐cell parameters a = b = 63.35, c = 125.90 Å, were obtained by vapour diffusion in hanging drops and the structure was refined to an Rfree of 16.7% at 1.5 Å resolution. The structure shows a CDR loop of the antibody on the GFP scaffold.