Almost all commercial proteins are purified using ammonium sulfate precipitation. Protein-polymer conjugates are synthesized from pure starting materials, and the struggle to separate conjugates from polymer, native protein, and from isomers has vexed scientists for decades. We have discovered that covalent polymer attachment has a transformational effect on protein solubility in salt solutions. Here, protein-polymer conjugates with a variety of polymers, grafting densities, and polymer lengths are generated using atom transfer radical polymerization. Charged polymers increase conjugate solubility in ammonium sulfate and completely prevent precipitation even at 100% saturation. Atomistic molecular dynamic simulations show the impact is driven by an anti-polyelectrolyte effect from zwitterionic polymers. Uncharged polymers exhibit polymer length-dependent decreased solubility. The differences in salting-out are then used to simply purify mixtures of conjugates and native proteins into single species. Increasing protein solubility in salt solutions through polymer conjugation could lead to many new applications of protein-polymer conjugates.

Resting state functional connectivity (RSFC) reflects the organization of functional networks in the brain. Functional networks measured during “resting”, or task-absent, state are correlated with cognitive function, and much development of these networks occurs between infancy and adulthood. However, RSFC research in the intermediate years (especially between ages 3 and 5 years) has been limited, mainly due to a paucity of child-appropriate neural measures and behavioral paradigms. This paper presents a new paradigm to measure RSFC in young children, utilizing functional near-infrared spectroscopy (fNIRS) and Freeplay, a simple behavioral setup designed to approximate resting state in children. In Experiment 1, we recorded fNIRS data from children aged 3-8 years and adults aged 18-21 years and examined feasibility and validity of our measure of RSFC, and compared measures across the two groups. In Experiment 2, we recorded longitudinal data at two points (approximately 3 months apart) from children aged 3-5 years, and examined reliability under a variety of measures. In both experiments, all children were able to complete testing and provide usable data, a significant improvement over fMRI-based RSFC measurement in children. Results suggest this paradigm is practical and has good construct validity and test-retest reliability, and may contribute towards increasing the availability of reliable data on resting state networks in early childhood. In particular, these are some of the first positive results on the feasibility of reliably measuring functional connectivity in children aged 3-5 years.