Figure 5. Thermal refolding efficiency determination of human VLs and VHs by SPR. (A), (B) Representative SPR sensorgrams for the binding of native and heat-denatured/cooled (refolded) human VLs to immobilized protein L. Data are from TRE experiments performed at 20 µM VL concentrations. VL concentrations used to construct each sensorgram set was 12.5, 18.8, 25, 37.5, 50, 75, and 100 nM for HVLP325 and 12.5, 20, 25, 30, 38, 50, and 75 nM for HVLP335. See Figure S3 and S4 for SPR data for all VLs and VHs. The KDns and KDrefs determined from “Native” and “Refolded” sensorgram pairs were used to determine TREs in (C). (C) TREs of wild-type VLs and VHs obtained under 4 µM and 20 µM domain concentration conditions in refolding experiments. Lines connect TREs for the same clones.