Recent advances in methods for quantifying the cell penetration of macromolecules

Area of Science:

• Biochemistry
• Cell Biology
• Drug Discovery

Background:

• Macromolecule therapeutics increasingly target intracellular components.
• Efficient delivery to intracellular compartments is essential for therapeutic efficacy.
• A gap often exists between biochemical binding affinity and cell-based activity.

Purpose of the Study:

• To review recent advancements in assays for quantifying macromolecule cellular uptake and cytosolic delivery.
• To address the need for robust methods to identify bottlenecks in intracellular drug delivery.
• To provide an overview of promising new assay technologies developed in the last 3-4 years.

Main Methods:

• Literature review of recent scientific publications (last 3-4 years).
• Focus on chemical biology approaches for measuring intracellular delivery.
• Analysis of assay advantages and disadvantages for quantifying cellular uptake and cytosolic delivery.

Main Results:

• Identification of a variety of novel assays for assessing macromolecule delivery.
• Discussion of the strengths and limitations of each assay type.
• Highlighting assays that accurately measure total cellular uptake and/or cytosolic delivery.

Conclusions:

• Robust assays are critical for understanding and optimizing macromolecule therapeutic delivery.
• Recent developments offer improved tools for chemical biologists and drug developers.
• Accurate measurement of intracellular delivery is key to bridging the gap between in vitro and in cellulo efficacy.