The continuous bed bioreactor systems have been used for the production of protein therapeutics, such as IgG, using immobilized enzyme in biopharmaceutical applications. We developed macroporous poly(hydroxyethyl methacrylate-co-glycidyl methacrylate) cryogel-based bioreactor matrix using sodium dodecyl sulfate as surfactans in the presence of ethylene glycol dimethacrylate as cross linking agent by bulk polymerization. The developed polyGMA immobilized bioreactor with papain enzyme was used for specific fragmentation of immunoglobulin G. The catalysis efficiency for immobilized enzyme were investigated in comparison with free enzyme. The immobilized papain displayed broad catalytic activity over a variety of conditions, with maximal activity around pH 7.0 and 70°C. The Michaelis-Menten kinetic constant (K_m), the maximum reaction velocity (V_max), and the catalytic efficiency (k_cat) for free enzyme were 0.1097 mg/mL, 29.9 mg/mL/min, and 92.01 1/min, respectively, whereas for immobilized enzyme, K_m, V_max, and k_cat values were 0.1078 mg/mL, 30.53 mg/mL/min, and 94.3 1/min, respectively. In a further step, after digestion, remarkable digestion products of bioreactor, Fab and Fc fragments, produced with immobilized papain bioreactors were analyzed in two ways by SDS-PAGE and reversed-phase HPLC; it was demonstrated that papain immobilized bioreactor successfully used for the digestion of human IgG with high activity. Therefore, the polyGMA cryogel immobilized with papain exhibited a very effective matrix for the bioreactor which can be considered as an alternative bioreactor matrix with great promise in biopharmaceutical applications.

连续床生物反应器系统已用于在生物制药应用中使用固定化酶生产蛋白质治疗剂，例如IgG。我们通过十二烷基硫酸钠作为表面活性剂，在乙二醇二甲基丙烯酸酯作为交联剂存在下，通过本体聚合反应，开发了基于大孔聚甲基丙烯酸羟乙酯-甲基丙烯酸缩水甘油酯共聚物的生物凝胶基质。将开发的木瓜蛋白酶固定化的polyGMA生物反应器用于免疫球蛋白G的特异性裂解。与游离酶比较，研究了固定化酶的催化效率。固定化的木瓜蛋​​白酶在各种条件下均显示出广泛的催化活性，在pH 7.0和70°C左右具有最大的活性。Michaelis-Menten动力学常数（K _m），游离酶的最大反应速度（V _max）和催化效率（k _cat）分别为0.1097 mg / mL，29.9 mg / mL / min和92.01 1 / min，而固定化酶的K _m，V _max和k _cat值分别为0.1078 mg / mL，30.53 mg / mL / min和94.3 1 / min。在进一步的步骤中，消化后，用固定化木瓜蛋白酶生物反应器产生的生物反应器，Fab和Fc片段的显着消化产物通过SDS-PAGE和反相HPLC进行了两种分析。已证明木瓜蛋白酶固定化生物反应器成功用于高活性人IgG的消化。因此，用木瓜蛋白酶固定化的polyGMA冷冻凝胶对生物反应器显示出非常有效的基质，可以将其视为替代生物反应器基质，在生物制药应用中具有广阔的前景。