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. 2011 Feb 4;30(6):1137–1148. doi: 10.1038/emboj.2011.21

Figure 1.

Figure 1

SPR analysis of the binding of TCR MS2-3C8 to MBP–DR4. (A) For kinetic measurements, TCR MS2-3C8 at concentrations of 2.9, 5.8, 11.5, 23 and 46 μM was injected over immobilized MBP–DR4 (320 RU). Sensograms were fitted to a 1:1 binding model to obtain on- and off-rates: kon=1.9 × 103±0.01 M−1 s−1 and koff=0.011±0.0003 s−1. KD was calculated as koff/kon. (B) For equilibrium measurements, TCR MS2-3C8 at concentrations of 1.2, 2.4, 4.7, 9.4 and 18.8 μM was injected over immobilized MBP–DR4 (400 RU). Inset shows the fitting curve for equilibrium binding that resulted in a KD of 5.0±0.1 μM. Equilibrium measurements for TCR MS2-3C8 mutants are also shown: (C) CDR1α T29A, (D) CDR3α K96A, (E) CDR2β E50A, (F) CDR2β T55A, (G) CDR3β S98A and (H) CDR3β N100A. For weakly binding mutants CDR3β S98A and CDR3β N100A, KDs were estimated at >300 and >100 μM, respectively.