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. 2012 Dec 10;7(12):e51397. doi: 10.1371/journal.pone.0051397

Figure 1. Expression and purfication of the Env183/A2 soluble T cell receptor.

Figure 1

(A) The gene encoding for TCR α chain residues 1–202 and TCR β chain residues 1–243 were separately cloned into pET28a vectors. A BirA recognition sequence was added to the C-terminus of the β chain. Cα Thr48Cys and Cβ Ser57Cys mutations were introduced to facilitate an inter-chain disulfide bond (red bar). (B) SDS PAGE analysis of two distinct fractions obtained after chromatographic purification of refolded TCR. (C) Gel shift analysis of the purified TCR shows >90% biotinylation. (D) Surface plasmon resonance analysis of refolded TCR monomers demonstrated functional binding to immobilized Env183/A2 with a KD of 0.6 µM, as determined with a steady-state model.