The ortho-phospho-tyrosine (P-Tyr) pseudoaffinity ligand was immobilized via ether linkage onto polyacrylamide-alginate (PAAm-Alg)-epoxy cryogels prepared according to two different approaches in order to explore their performance in the immunoglobulin G (IgG) purification from human serum. In the first approach, the P-Tyr was attached to cryogel prepared by cryocopolymerization of acrylamide and alginate with allyl glycidyl ether (AGE) as functional comonomer, and methylenebisacrylamide and Ca(II) as crosslinkers, obtaining the PAAm-Alg-AGE-P-Tyr. In the second approach, the PAAm-Alg was synthesized under the same conditions, but without AGE, and the P-Tyr was coupled to epichlorohydrin (ECH)-activated cryogel, obtaining the PAAm-Alg-ECH-P-Tyr. Both pseudoaffinity cryogels were characterized by scanning electron microscopy, swelling tests, porosity, ligand density, and flow dynamics. The human IgG differently interacted with the PAAm-Alg-ECH-P-Tyr and PAAm-Alg-AGE-P-Tyr cryogels, depending on the pH and adsorption buffer system used. The selectivity analyzed by electrophoretic profiles was similar for both cryogels, but PAAm-Alg-ECH-P-Tyr achieved higher IgG adsorption capacity (dynamic capacity of 12.62 mg of IgG/mL of cryogel). The IgG purity assayed by ELISA was 95%. The maximum IgG adsorption capacity and dissociation constant of the PAAm-Alg-ECH-P-Tyr, determined by Langmuir isotherm, were found to be 91.75 mg IgG/g of dry cryogel and 4.60 × 10⁻⁶ mol/L at pH 6.0 from aqueous solutions. The PAAm-Alg-AGE-P-Tyr showed potential to purify the Fab fragments from papain-digested human IgG solution at pH 7.0. Fab fragments were separated from Fc fragments (but with uncleaved IgG) in eluted fractions (analyzed by the Western blot technique), with yield of 82% and purity of 95% (determined by radial immunodiffusion).

在邻通过醚键将-磷酸-酪氨酸（P-Tyr）假亲和配体固定在根据两种不同方法制备的聚丙烯酰胺-海藻酸酯（PAAm-Alg）-环氧冰冻剂上，以研究其在纯化人免疫球蛋白G（IgG）中的性能血清。在第一种方法中，将P-Tyr连接到通过丙烯酰胺和藻酸盐与烯丙基缩水甘油醚（AGE）作为功能性共聚单体，亚甲基双丙烯酰胺和Ca（II）作为交联剂的低温共聚制备的冷冻凝胶上，获得PAAm-Alg-AGE-P蒂尔 在第二种方法中，在相同条件下但没有AGE合成PAAm-Alg，并将P-Tyr与环氧氯丙烷（ECH）活化的冷冻凝胶偶联，得到PAAm-Alg-ECH-P-Tyr。两种假亲和力凝胶均通过扫描电子显微镜，溶胀测试，孔隙率，配体密度和流动动力学。根据使用的pH值和吸附缓冲液系统，人IgG与PAAm-Alg-ECH-P-Tyr和PAAm-Alg-AGE-P-Tyr低温凝胶的相互作用不同。通过电泳图谱分析的选择性对于两种冷冻凝胶都是相似的，但是PAAm-Alg-ECH-P-Tyr获得了更高的IgG吸附容量（动态容量为12.62 mg IgG / mL冷冻凝胶）。通过ELISA测定的IgG纯度为95％。通过Langmuir等温线测定，发现PAAm-Alg-ECH-P-Tyr的最大IgG吸附能力和解离常数为91.75 mg IgG / g干冰凝胶和4.60×10 通过电泳图谱分析的选择性对于两种冷冻凝胶都是相似的，但是PAAm-Alg-ECH-P-Tyr获得了更高的IgG吸附容量（动态容量为12.62 mg IgG / mL冷冻凝胶）。通过ELISA测定的IgG纯度为95％。通过Langmuir等温线测定，发现PAAm-Alg-ECH-P-Tyr的最大IgG吸附能力和解离常数为91.75 mg IgG / g干冰凝胶和4.60×10 通过电泳图谱分析的选择性对于两种冷冻凝胶都是相似的，但是PAAm-Alg-ECH-P-Tyr获得了更高的IgG吸附容量（动态容量为12.62 mg IgG / mL冷冻凝胶）。通过ELISA测定的IgG纯度为95％。通过Langmuir等温线测定，发现PAAm-Alg-ECH-P-Tyr的最大IgG吸附能力和解离常数为91.75 mg IgG / g干冰凝胶和4.60×10来自水溶液的pH值为^-6 mol / L。PAAm-Alg-AGE-P-Tyr在pH 7.0时显示出从木瓜蛋白酶消化的人IgG溶液中纯化Fab片段的潜力。Fab片段与Fc片段（但有未切割的IgG）从洗脱片段中分离（通过Western blot技术分析），产率为82％，纯度为95％（通过放射免疫扩散测定）。