Journal: Biomacromolecules

Article Title: Efficient Self-Immolative RAFT End Group Modification for Macromolecular Immunodrug Delivery

doi: 10.1021/acs.biomac.3c00239

Figure Lengend Snippet: Characterization and application of RAFT polymer–immunodrug conjugates demonstrating the superiority of the in situ -introduced self-immolative linker strategy. (A) Post-polymerization modification of fluorescently labeled pDMA (average DP = 40) without active component (top), IMDQ bridged via the self-immolative motif (middle) and IMDQ attached via thioether (bottom). (B) MALDI-ToF MS data of respective DP with highest relative intensity and its corresponding simulated isotope pattern in red. (C) UV–vis spectra of post-polymerization-modified polymers exhibiting the characteristic IMDQ absorbance at 322 nm. (D) Illustration of the in vitro reporter cell assay. Cells were incubated with polymer samples on a 96-well plate for 48 h, when the polymer samples can get intracellularly processed and QUANTI-Blue assay can be performed to verify enhanced receptor activity for the SIL conjugate. Color intensity, reflecting induced phosphatase activity, rises from increasing IMDQ activity. (E) Results of the assay readout. The data of the self-immolative linked IMDQ (red) were shifted toward free IMDQ (black) in comparison to thioether-linked IMDQ (red, dotted). Polymer without active compound exhibited no activity at all (red, dashed) (illustration of 96-well plates was adapted from BioRender.com).

Article Snippet: QUANTI-Blue secreted alkaline phosphatase and MTT assay readouts were performed on a Spark 20M multimode microplate reader from TecanTrading AG (Mannedorf, Switzerland).

Techniques: In Situ, Modification, Labeling, In Vitro, Incubation, Activity Assay