Figure 4: Construction of homogenous and functional antibody-conjugates. | Nature Communications

Figure 4: Construction of homogenous and functional antibody-conjugates.

From: Stoichiometric and irreversible cysteine-selective protein modification using carbonylacrylic reagents

Figure 4

(a) Scheme for chemoselective and equimolar bioconjugation of Trastuzumab with 2c and 7. (b,c) Electrospray ionization–MS spectra of the light-chain of Trastuzumab (b) before and (c) after conjugation with 2c. (d) BLI curves and fit for Trastuzumab-2c and HER2. KD constants derived from BLI experiments for non-modified Trastuzumab and conjugated Trastuzumab-2c. (e,f) Analysis of specificity of the Trastuzumab-7 towards HER2 by flow-cytometry. (e) Percentage of FITC-positive single cells, after treatment with fluorescent Trastuzumab-7 or non-fluorescent Trastuzumab. (f) Superposition of contour plots of side-scatter detection versus FITC-equivalent fluorescence intensity, in SKBR3 cells (expressing high levels of her2/c-erb-2) and in HepG2 cells (expressing low levels of her2/c-erb-2). Controls were treated with non-conjugated Trastuzumab and samples were treated with increasing concentrations of Trastuzumab-7 (10, 50 and 150 nM). RT, room temperature.

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