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. Author manuscript; available in PMC: 2020 May 16.
Published in final edited form as: Cell Chem Biol. 2019 Feb 28;26(5):634–644.e3. doi: 10.1016/j.chembiol.2019.02.001

Figure 1-. PEGylated aptamers are recognized by anti-PEG IgGs in patient plasma.

Figure 1-

(a) Schematic representation of the PEGylated RNA aptamer RB006.

(b) Indirect ELISA demonstrates that plasma from anti-PEG IgG positive patients (blue) exclusively detects PEGylated compounds including the PEGylated RNA aptamers RB006 and Macugen and the PEGylated proteins ADAgen (PEGylated adenosine deaminase). However, patient plasma containing anti-PEG antibodies did not detect the non-PEGylated controls, including a non-PEGylated version of ADAgen, termed ADA, and a non-PEGylated version of RB006, termed RB005. Additionally, patient plasma that is negative for anti-PEG IgGs (grey) did not detect any of the PEGylated compounds. Data represent the mean ± SD of technical triplicates. *** denotes p-value < 0.001, using a t-test against (−) PEG plasma.

(c) Competition ELISA demonstrates that patient-derived anti-PEG antibodies bind with more affinity to PEGylated verses unPEGylated compounds, as shown by a decrease in absorbance with increasing concentrations of either the PEGylated aptamer RB006 (blue) or the PEGylated protein ADAgen (orange). No change was seen with the unPEGylated controls RB005 (red) and ADA (purple). Data represents the mean ± SD of technical triplicates. ** denotes p-value<0.01, *** denotes p-value < 0.001, using t-test verses unPEGylated compounds.