Temperature-responsive assembly/disassembly of nanostructures around body temperature is a valuable means of controlling encapsulation/release of therapeutic cargo. Modulation of the transition temperature allows for application-specific optimisation of cargo loading and delivery, which are critical considerations in the drug delivery field. The synthesised P(NIPAMco-NCPAM)-based triblock copolymers were designed to exhibit increasing transition temperatures. DLS was used to confirm successful modulation of the self-assembly temperature for the P(NIPAM-co-NCPAM)-based triblock copolymers containing increasing amounts of the cyclopropyl-based repeating unit (NCAPM from 0 to 60%) (5a4/5c1-3) (Figure ). By dissolving these copolymers in PBS at 25 °C and increasing the temperature to 60 °C (above the LCST), a clear increase in the turbidity of the solution could be observed by eye and by light scattering (derived count rate, kcps) recorded using DLS, which suggests a coilto-globule transition (Figure ). Since larger structures scatter more light than dissolved copolymers, the self-assembly of the copolymers could be followed by monitoring the scattering intensity. The inflection points of each curve represent the Tcp of the corresponding copolymers. DLS measurements at temperatures below/above the LCST indicate formation of stable and well-defined nanostructures when heated above the LCST (Figure ), whilst bigger structures were formed with increasing amounts of NCPAM (mean hydrodynamic diameters between 74 and 295 nm). Due to the formation of large structures/aggregates that led to some precipitation for 5c3 when heated/cooled slowly (1 °C temperature ramp with 5 min equilibration time), the intensity curves are not smooth (Figure ) and no accurate Tcp can be given for this copolymer. However, it appears to be between 45 to 50 °C when estimating based on both curves (up/down, heating and cooling). The dependence of particle size on the heating rate during the self-assembly process was also confirmed for two copolymers that form smaller nanoparticles in general (5a4 and 5c1, Figure ). The final assembled particles tended to be larger when subjected to slower heating. The cyclopropyl-dependent size increase might be caused by the higher rigidity and lower rotational freedom of the cyclopropyl group when compared to the isopropyl group, which might affect the packing parameters to yield larger structures. The self-assembly temperature of the resulting triblock copolymers was successfully tuned from 33 °C, corresponding to the Tcp of the 5a4 copolymer with 0% NCPAM content, to a Tcp of about 47 °C for the 5c3 copolymer with 60% NCPAM content. It can be concluded that by increasing the number of cyclopropyl units in the copolymers, the self-assembly temperature increases proportionally. However, when the NCPAM content exceeds 50%, the stability of the resulting self-assembled structures is compromised due to the formation of large particles/aggregates, which can lead to sedimentation (Figure ). The self-assembly process of PNIPAM-and P(NIPAM-co-NCPAM)-based copolymers can be reversed by cooling the solution below the transition temperature (down to 25 °C) causing complete nanogel dissolution (Figure ). The globuleto-coil transition is slower than the coil-to-globule transition, which is indicated by the slight hysteretic behaviour. According to this initial DLS analysis, the selected block copolymers (5a4/5c1-3) revealed an LCST-type, reversible thermo-responsive behaviour that yielded nanogels at temperatures above their LCST. We also tested all the other copolymers from the separate PNIPAM (5a1 to 5a5) and PNCPAM (5b1 to 5b5) series for the formation of nanogels at temperatures above the LCST (Table ). All copolymers except 5b1 assembled into nanogels, however, no clear trends regarding polymer lengths and corresponding nanostructure sizes could be found.