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. 2021 May 25;12:3142. doi: 10.1038/s41467-021-23274-x

Fig. 1. NI301A binds to wild-type and mutant ATTR fibrils in vitro, with absent binding to native TTR tetramers.

Fig. 1

a Characterization of ATTRwt fibrils prepared in vitro: fibrils were visualized by electron microscopy after 72 h and bound the amyloid indicator dye thioflavin T. b NI301A binding to coated ATTRwt and ATTRv fibrils by ELISA (symbols: mean of duplicates, lines: data fitting). c Sandwich ELISA using NI301A for capture and detection to measure in-solution binding to ATTR and TTR tetramers (mean of duplicates). d SPR analysis of NI301A-binding kinetics to ATTRwt oligomers and native TTRwt tetramers in solution. e BLI analysis of NI301A-binding kinetics to ATTR-V30M and ATTR-V122I oligomers, and native TTRv tetramers in solution. For d and e, data (gray to black) were fitted with a 1 : 1 binding model (red lines). f Dot blot analysis of NI301A binding to native tetramers or amyloid aggregates of TTRwt and TTRv proteins. A polyclonal, pan-TTR antibody (Dako A0002) was used for comparison. g Analysis of TTR-V30M aggregation time course by semi-native SDS-PAGE followed by western blotting with antibodies NI301A, 39-44, and Dako A0002. Positions of ATTR monomers (1), dimers (2), trimers (3), tetramers (4), and aggregates of high molecular weight (HMW) are marked. h Immunoprecipitation with antibodies NI301A, Dako A0002, and human IgG1 isotype control of native TTR tetramers (tetr.TTR) or patient-derived ATTR fibrils (ATTR fibr.), and serial dilutions of human plasma from healthy donors.