6WZN

Crystal Structure of a Fluorescent Single Chain Fv Chimera

Summary for 6WZN

• Entry DOI: 10.2210/pdb6wzn/pdb
• Descriptor: Fluorescent Single Chain Fv Chimera, GLYCEROL (3 entities in total)
• Functional Keywords: thermal-stable fluorescent protein, single chain fv, fusion, engineered protein, antibody, fluorescent protein
• Biological source: synthetic construct
• Total number of polymer chains: 1
• Total formula weight: 51083.91

Authors

Close, D.,Velappan, N.,Hung, L.W.,Naranjo, L.,Hemez, C.,DeVore, N.,McCullough, D.,Lillo, A.M.,Waldo, G.,Bradbury, A.R.M. (deposition date: 2020-05-14, release date: 2021-01-27, Last modification date: 2024-10-30)

Primary citation

Velappan, N.,Close, D.,Hung, L.W.,Naranjo, L.,Hemez, C.,DeVore, N.,McCullough, D.K.,Lillo, A.M.,Waldo, G.S.,Bradbury, A.R.M.
Construction, characterization and crystal structure of a fluorescent single-chain Fv chimera.
Protein Eng.Des.Sel., 34:-, 2021

PubMed Abstract: In vitro display technologies based on phage and yeast have a successful history of selecting single-chain variable fragment (scFv) antibodies against various targets. However, single-chain antibodies are often unstable and poorly expressed in Escherichia coli. Here, we explore the feasibility of converting scFv antibodies to an intrinsically fluorescent format by inserting the monomeric, stable fluorescent protein named thermal green, between the light- and heavy-chain variable regions. Our results show that the scTGP format maintains the affinity and specificity of the antibodies, improves expression levels, allows one-step fluorescent assay for detection of binding and is a suitable reagent for epitope binning. We also report the crystal structure of an scTGP construct that recognizes phosphorylated tyrosine on FcεR1 receptor of the allergy pathway.

PubMed: 33586761
DOI: 10.1093/protein/gzaa029

Experimental method

X-RAY DIFFRACTION (2.5 Å)