Continuous preparation of bicelles using hydrodynamic focusing method for bicelle to vesicle transition

Bicelle is one of the most stable phospholipid assemblies, which has tremendous applications in the research areas for drug delivery or structural studies of membrane proteins owing to its bio-membrane mimicking characteristics and high thermal stability. However, the conventional preparation method for bicelle demands complicated manufacturing processes and a long time so that the continuous synthesis method of bicelle using microfluidic chip has been playing an important role to expand its feasibility. We verified the general availability of hydrodynamic focusing method with microfluidic chip for bicelle synthesis using various kinds of lipids which have a phase transition temperature ranged from - 2 to 41 degrees C. Bicelle can be formed only when the inside temperature of microfluidic chip was over the phase transition temperature. Moreover, the concentration condition for bicelle formation varied depending on the lipids. Furthermore, the transition process characteristics from bicelle to vesicle were analyzed by effective q-value, mixing time and dilution condition. We verified that the size of transition vesicles was controlled according to the effective q-value, mixing time, and temperature.

Automated summary

Bicelle, as a stable phospholipid assembly with bio-membrane mimicking characteristics, holds great potential in drug delivery and membrane protein structural studies. The continuous synthesis method using a microfluidic chip has been crucial in enhancing its feasibility. Different types of lipids with varying phase transition temperatures were successfully used in synthesizing Bicelle via hydrodynamic focusing method in a microfluidic chip, with formation being temperature-dependent and requiring specific concentration conditions. The transition process characteristics from Bicelle to vesicles were analyzed, showing that the size of transition vesicles can be controlled by factors such as effective q-value, mixing time, and temperature.