LNPs and nanocarriers

Question 1

LNP life cycle

Answer 1

1. Formulation - Microfluidic mixing
2. Dialysis - Remove ethanol, neutralize ionizable lipid
3. Peg-shredding coronation - Increases target affinity
4. Endocytosis - Entering of the cell
5. Endosomal escape –> protein production
6. Recycling
7. Cell to cell transfer?

Question 2

EVs

Answer 2

Extracellular vesicles. Occurs naturally in the body so very good nano carriers. More complex and not as easy to manufacture as LNPs

Question 3

DLS

Answer 3

Dynamic light scattering. Checks light scattered by particles during brownian motion. Large particles move slower –> less fluctuation in scattering. Gives information about NP size

Question 4

NTA

Answer 4

Nanoparticle Tracking Analysis. Brownian motion of INDIVIDUAL particles (red line follows each particle). Can handle more dilute samples than DLS. Concentration and size information.

Question 5

Cryo-TEM

Answer 5

Cryo Transition Electron Microscope. I assume the particles are easy to identify is cause they are very cold so there is almost no brownian motion?

1. Sample is placed on a metal grid
2. Grid gets frozen by liquid nitrogen and is then measured using TEM

Question 6

DSC

Answer 6

Differential scanning calorimetry. Measures specific heat capacity. Gives thermoscientific data such as phase transistion temperatures. I.e if you add cholesterol to DSPC the phase transition is less dramatic.

Question 7

Small angle scattering

Answer 7

Using X-rays or neutrons to probe structrual arrangement of matter. Can be used to study particle morphology and lipid phase behaviour.