Protein crystallization has been shown as a viable alternative to conventional protein purification methods. However, the stochastic and spontaneous nature of the primary nucleation process makes protein crystallization difficult to control. An approach often used in the crystallization of small organic and inorganic molecules to improve the reproducibility of the process and better control the crystallization outcome is seeding. This study presents for the first time the investigation of seeding on batch protein crystallization in a meso oscillatory flow reactor (meso-OFR). Seeded and unseeded lysozyme batch crystallization experiments were carried out, where the influence of the initial lysozyme concentration (initial supersaturation) and the seed quantity on the onset of nucleation, crystal properties and yield was assessed. No major influence of the seed quantity on the product crystal size was verified for the initial lysozyme concentration of 20 mg·mL⁻¹ (below the secondary nucleation threshold - SNT), while for the initial lysozyme concentration of 25 mg·mL⁻¹ (above the SNT) the onset of nucleation and mean crystal size significantly decrease with increasing seed quantity. Further, crystal yield increases with both the initial lysozyme concentration and seed quantity. Overall, the results highlight seeding as a promising technique to improve control over the crystallization time and product crystal size.

蛋白质结晶已被证明是传统蛋白质纯化方法的可行替代方法。然而，初级成核过程的随机性和自发性使得蛋白质结晶难以控制。一种常用于有机和无机小分子结晶以提高工艺再现性和更好地控制结晶结果的方法是晶种。本研究首次介绍了在中观振荡流反应器 (meso-OFR) 中对分批蛋白质结晶进行播种的研究。进行了有种子和无种子的溶菌酶批量结晶实验，评估了初始溶菌酶浓度（初始过饱和度）和种子数量对成核开始、晶体性质和产量的影响。^-1（低于二次成核阈值 - SNT），而对于 25 mg·mL ^-1的初始溶菌酶浓度（高于 SNT），成核开始和平均晶体尺寸随着晶种数量的增加而显着降低。此外，晶体产量随着初始溶菌酶浓度和种子数量的增加而增加。总体而言，结果强调了晶种作为一种有前途的技术，可以改善对结晶时间和产品晶体尺寸的控制。