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. Author manuscript; available in PMC: 2020 Jan 1.
Published in final edited form as: Nat Chem. 2018 Nov 5;11(1):78–85. doi: 10.1038/s41557-018-0154-0

Figure 3. Conjugation of vancomycin to affibody generates conjugates with retained structure and binding affinity.

Figure 3.

(a) Conjugation of affibody to vancomycin. The affibody variant 27 containing activated selenocysteine was generated in situ from variant 25 (Supplementary Section 23). (b) Deconvolved mass spectrum of purified affibody-vancomycin conjugate. (c) Far-UV Circular dichroism (CD) spectrum of affibody, affibody-selenocysteine variant 25, and affibody-vancomycin conjugate 27. The CD spectrum of 27 was obtained by subtracting the measured spectra with that of vancomycin only (Supplementary Section 24). (d) Sensorgrams from BioLayer interferometry analysis of affibody-vancomycin conjugate binding to trastuzumab antibody. Retained binding was observed compared to the native affibody (KD = 2.0 ± 0.2 nM; the final KD is the average of the KD obtained from three different concentrations, the error is standard error.)