Impact of poly(ethylene glycol) functionalized lipids on ordering and fluidity of colloid supported lipid bilayers

Colloid supported lipid bilayers (CSLBs) are highly appealing building blocks for functional colloids. In this contribution, we critically evaluate the impact on lipid ordering and CSLB fluidity of inserted additives. We focus on poly(ethylene glycol) (PEG) bearing lipids, which are commonly introduced to promote colloidal stability. We investigate whether their effect on the CSLB is related to the incorporated amount and chemical nature of the lipid anchor. To this end, CSLBs were prepared from lipids with a low or high melting temperature (T_m), DOPC, and DPPC, respectively. Samples were supplemented with either 0, 5 or 10 mol% of either a low or high T_m PEGylated lipid, DOPE-PEG₂₀₀₀ or DSPE-PEG₂₀₀₀, respectively. Lipid ordering was probed via differential scanning calorimetry and fluidity by fluorescence recovery after photobleaching. We find that up to 5 mol% of either PEGylated lipids could be incorporated into both membranes without any pronounced effects. However, the fluorescence recovery of the liquid-like DOPC membrane was markedly decelerated upon incorporating 10 mol% of either PEGylated lipids, whilst insertion of the anchoring lipids (DOPE and DSPE without PEG₂₀₀₀) had no detectable impact. Therefore, we conclude that the amount of incorporated PEG stabilizer, not the chemical nature of the lipid anchor, should be tuned carefully to achieve sufficient colloidal stability without compromising the membrane dynamics. These findings offer guidance for the experimental design of studies using CSLBs, such as those focusing on the consequences of intra- and inter-particle inhomogeneities for multivalent binding and the impact of additive mobility on superselectivity.