Supplementary Figure 15: Mass spectra of anti-RCA scFvs.
From: Custom selenoprotein production enabled by laboratory evolution of recoded bacterial strains
(a) Mass spectrum of intact wild-type anti-RCA scFv. (b) Deconvoluted mass spectrum of wild-type anti-RCA scFv. (c) Experimentally determined monoisotopic mass (27002.21 Da) matches the theoretical mass (27002.17 Da) of wild-type anti-RCA scFv containing a two disulfide bonds. (d) UVPD mass spectrum of the 20+ charge state. (e) The UVPD fragment map shows a lack of detectable fragments between C23 and C97 in the VH domain, and C159 and C224 in the VL domain, which confirms correct formation of both disulfide bonds. (f) Mass spectrum of intact seleno-anti-RCA. (g) Deconvoluted mass spectrum of seleno-anti-RCA. (h) Experimentally determined monoisotopic mass (27193.96 Da) matches the theoretical mass (27193.94 Da) of seleno-anti-RCA containing two diselenide bonds. The experimental mass has been adjusted to account for deconvolution error arising from the unusual isotope distribution of selenium. (i) UVPD mass spectrum of the 20+ charge state. (j) The UVPD fragment map shows a lack of detectable fragments between U23 and U97 in the VH domain, and U159 and U224 in the VL domain, which confirms correct formation of both diselenide bonds. All spectra are representative from a minimum of two technical replicates.
