The present work deals with the development of novel affinity monolith with reversible protein ligands for protein chromatography. As for the formation of reversible ligands, Concanavalin A (Con A) is chelated with Cu(II)-iminodiacetic acid (IDA) immobilized macroporous cellulose monolith (MCM) for glycoprotein adsorption. The reversible immobilization is realized by Cu ions, which bridge affinity ligands and support by strong chelation interaction. The fabrication process of reversible Con A immobilized adsorbent is studied, especially with regards to the effect of synthesis conditions on the ligands immobilization. The adsorption behavior is then evaluated to elucidate the potential of Con A-Cu(II)-IDA-MCM for protein chromatography. It reveals that the static adsorption capacity and dissociation constant of glucose oxidase (GOD) on Con A-Cu(II)-IDA-MCM are determined to be 17.4 ± 0.6 mg mL⁻¹ and 0.055 ± 0.011 mg mL⁻¹ by Langmuir model. With frontal analysis, the dynamic binding capacity of GOD at 10% breakthrough point is about 11.4 ± 1.0 mg mL⁻¹ and changes less with an increase of flow velocity from 0.2 to 1.0 mL min⁻¹. Moreover, Con A-Cu(II)-IDA-MCM displays weak nonspecific adsorption for the impurities and is able to successfully enrich glycoprotein ovalbumin (OVA) from diluted chicken egg white. In addition, Con A-Cu(II)-IDA-MCM exhibits excellent stability by the repeated adsorption/desorption operations. By taking these advantages of high adsorption capacity, excellent specificity and structure stability, the prepared affinity adsorbent of Con A-Cu(II)-IDA-MCM has great potential for high performance protein chromatography.

本工作涉及具有可逆蛋白质配体的新型亲和整体料的开发，用于蛋白质色谱。至于可逆配体的形成，将伴刀豆球蛋白A（Con A）与固定有铜（II）-亚氨基二乙酸（IDA）的大孔纤维素整料（MCM）螯合，以吸附糖蛋白。可逆的固定化是通过Cu离子实现的，Cu离子通过强大的螯合相互作用将亲和配体和载体连接在一起。研究了可逆Con A固定化吸附剂的制备过程，特别是关于合成条件对配体固定化的影响。然后评估吸附行为，以阐明Con A-Cu（II）-IDA-MCM在蛋白质色谱中的潜力。Langmuir模型的^-1和0.055±0.011 mg mL ^-1。通过正面分析，GOD在10％突破点的动态结合能力约为11.4±1.0 mg mL ^-1，并且随着流速从0.2到1.0 mL min ^-1的增加而变化较小。此外，Con A-Cu（II）-IDA-MCM对杂质显示出较弱的非特异性吸附，并能够成功地从稀释的​​鸡蛋白中富集糖蛋白卵清蛋白（OVA）。另外，Con A-Cu（II）-IDA-MCM通过重复的吸附/解吸操作表现出优异的稳定性。利用这些高吸附容量，优异的特异性和结构稳定性的优点，制备的Con A-Cu（II）-IDA-MCM亲和吸附剂在高效蛋白质色谱中具有巨大的潜力。