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Delivery of impermeable peptides to enable early pharmacological validation

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Who this is for:

  • Early stage drug-development teams working on peptides, macrocycles, and other challenging molecules

Opportunities

  • Direct intracellular delivery provides cellular pharmacological validation for peptides and macrocycles before additional chemistry investment, supporting Direct-to-Biology assays
  • With this information, you can save time and resources by only investing in scaffolds that have desirable target engagement and biological effect
  • Enables decoupling of optimization for permeability from optimization for potency/activity. Providing medicinal chemists with more valuable insights during their campaigns.

Results Obtained

  • Robust delivery of cyclic and non-cyclic peptides to HEK293 and HeLa cells for drug screening

How We Did It

Portal uses mechanoporation to deliver virtually any cargo to diverse cell types while maintaining cell health. This enables intracellular delivery of impermeable molecules without the complications of electroporation or other delivery methods. Learn more about mechanoporation here! 

Macrocycle Delivery to HeLa cells Shows Robust Dose Response

When boosted with a FITC labelled peptide macrocycle, HeLa cells showed a robust dose response inside the cell as demonstrated by the right shift of the histogram and good viability (Fig 1 & 2). The experiment was executed in a 96 well plate, supporting high throughput screening.  Flow cytometry was conducted post-boost to assess viability and efficiency of macrocycle delivery into boosted cells.

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Fig 1: HeLa cells were boosted then manually added into wells of a 96-well plate pre-loaded with FITC-labeled macrocycle at 0-0.2 mg/ml. Flow cytometry was conducted post-boost to assess viability.

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Fig 2: HeLa cells were boosted then manually added into wells of a 96-well plate pre-loaded with FITC-labeled macrocycle at 0-0.2 mg/ml. Flow cytometry was conducted post-boost to assess efficiency of macrocycle delivery into boosted cells

Efficient Delivery of Labeled Macrocycle to HEK293 Cells

Similar results were obtained for HEK293 cells with efficient delivery shown using a FITC labelled macrocycle. Viability and macrocycle delivery to the untreated and boosted samples were measured by flow cytometry (Fig. 3) and fluorescence microscopy (Fig. 4).

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Fig 3: HEK293 cells were mechanoporated with a FITC-labeled macrocycle. Viability and macrocycle delivery to the untreated and boosted samples were measured by flow cytometry.
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Fig 4:  HEK293 cells were mechanoporated with a FITC-labeled macrocycle. Viability and macrocycle delivery to the untreated and boosted samples were measured by fluorescence microscopy. Scale bar = 300 μm.

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