Fluorinated and Delaminated Ti3C2 MXene as Heterogeneous Nucleants to Assist Lysozyme Crystallization

Abstract Protein crystallization is vital for drug purification and protein structure analysis. However, spontaneous protein nucleation features high randomness and low efficiency, and tends to trigger disordered protein aggregation, which greatly restricts its practical applications. Accordingly, efficient regulation of protein crystallization can enhance crystallization efficiency, shorten experimental duration, reduce sample consumption, and further facilitate protein structure determination and biopharmaceutical research. This study adopted two types of two-dimensional materials, namely fluorinated Ti3C2 MXene (f-Ti3C2) and delaminated Ti3C2 MXene (d-Ti3C2), as heterogeneous nucleants to investigate their effects on lysozyme crystallization. The experiment was carried out by introducing different concentrations of f-Ti3C2 and d-Ti3C2 into the lysozyme solution system, and continuously monitoring and recording the process of protein nucleation and crystallization. The results suggest that at low protein concentrations, f-Ti3C2 and d-Ti3C2, acting as heterogeneous nucleants, can effectively enhance protein crystallization efficiency and address the difficulty in crystallizing proteins at low concentrations. Furthermore, both materials effectively regulate the average number and size of protein crystals. Between them, delaminated d-Ti3C2 exhibits superior performance in modulating crystallization success rate and crystal quantity. This study not only extends the modified two-dimensional Ti3C2 MXene to the field of protein crystallization and opens up new application avenues for such materials, but also diversifies the range of novel heterogeneous nucleants available for protein crystallization and effectively optimizes and regulates protein nucleation and crystallization.