Recording the data necessary to assess the kinetics of a reaction can be labor-intensive. In this technology brief, we show a method to automate this task by utilizing parts of an ÄKTApurifier chromatography system to automatically take samples from a reaction vessel at predefined time intervals and place them in 96-well plates and also enable correlating the samples with in-line spectral data of the reaction solution. Automatic sampling can reduce experimental bottlenecks by enabling overnight reactions or a higher degree of parallelization. To demonstrate the feasibility of the method, we performed batch-PEGylation of lysozyme with varying conditions by changing the molar excess of the PEG reagent. We used analytical cation-exchange chromatography to analyze the samples taken during the batch reaction, determining the concentrations of the individual species present at each time step. Subsequently, we fitted a kinetic model on these data. Fitting the model to four different reaction conditions simultaneously yielded a regression coefficient of R² = 0.871.

记录评估反应动力学所需的数据可能是劳动密集型的。在本技术简报中，我们展示了一种通过利用 ÄKTApurifier 色谱系统的一部分自动从反应容器中以预定义的时间间隔采集样品并将其放置在 96 孔板中，并使样品与内反应溶液的线光谱数据。自动采样可以通过启用过夜反应或更高程度的并行化来减少实验瓶颈。为了证明该方法的可行性，我们通过改变 PEG 试剂的摩尔过量，在不同条件下对溶菌酶进行了批量聚乙二醇化。我们使用分析型阳离子交换色谱法来分析批反应期间采集的样品，确定每个时间步长存在的单个物种的浓度。随后，我们对这些数据建立了动力学模型。将模型同时拟合到四种不同的反应条件，得到的回归系数为R ² = 0.871。