The partitioning and purification of lectins from the crude extract of Cratylia mollis seeds (Cramoll 1,4) was investigated in aqueous two-phase systems (ATPS). A factorial design model (2⁴) was used to evaluate the influence of polyethylene glycol (PEG) molar mass (1500–8000 g/mol), PEG concentration (12.5–17.5% w/w), phosphate (10–15% w/w) concentration, and pH (6–8) on the differential partitioning, purification factor, and yield of the lectin. Polymer and salt concentration were the most important variables affecting partition of lectin and used to find optimum purification factor by experimental Box–Behnken design together with the response surface methodology (RSM). ATPS showed best conditions composed by 13.9% PEG1500, 15.3% phosphate buffer at pH 6, which ensured purification factor of 4.70. Sodium dodecyl sulfate–polyacrylamide gel electrophoresis showed a single band of protein with 26.1 kDa. Furthermore, results demonstrated a thermostable lectin presenting activity until 60 °C and lost hemagglutinating activity at 80 °C. According to the obtained data it can be inferred that the ATPS optimization using RSM approach can be applied for recovery and purification of lectins.

在含水两相系统（ATPS）中研究了从Cratylia mollis种子（Cramoll 1,4）的粗提物中提取的凝集素的分配和纯化。析因设计模型（2 ⁴）用于评估聚乙二醇（PEG）摩尔质量（1500–8000 g / mol），PEG浓度（12.5–17.5％w / w），磷酸盐（10–15％w / w）浓度和pH的影响（6–8）关于凝集素的差异分配，纯化因子和产率。聚合物和盐浓度是影响凝集素分配的最重要变量，可通过实验Box–Behnken设计和响应面方法（RSM）来找到最佳纯化因子。ATPS表现出最佳条件，该条件由pH值为6的13.9％PEG1500、15.3％磷酸盐缓冲液组成，可确保纯化系数为4.70。十二烷基硫酸钠-聚丙烯酰胺凝胶电泳显示一条蛋白带，分子量为26.1 kDa。此外，结果表明直到60°C的热稳定凝集素呈现活动，并在80°C失去血凝活性。