Development of histidine-tagged cyclic peptide functionalized monolithic material for the affinity purification of antibodies in biological matrices
Development of histidine-tagged cyclic peptide functionalized monolithic material for the affinity purification of antibodies in biological matrices
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开发组氨酸标记的环肽功能化整体材料，用于生物基质中抗体的亲和纯化
DOI：
10.1016/j.chroma.2020.461707
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发表时间：
2021
影响因子：
4.1
通讯作者：
Jiang zhengjin
中科院分区：
文献类型：
--
作者：
Xu rongrong;Lu li;Sun lingjue;Liu xiao;Lei yutian;Huang shengfeng;Huang hao;Crommen Jacques;Han Hai;Wang Qiqin;Jiang zhengjin
随着单抗在治疗中应用的迅速增加，学术和工业用单抗生产和纯化技术的发展势在必行。本论文采用金属离子络合的方法合成了组氨酸标记的环肽(HHHHHGSGSDC*AWHLGELVWC*T，其二硫键半胱氨酸用星号表示，命名为HT25-环肽)功能化整体材料。所得材料具有合适的亲和力和多肽配体密度(每毫升材料中含有13.8 mg多肽配体)、良好的孔隙率(67.1%)、可接受的比表面积(52.95m2/g)和大量的大孔(4.13μm)。此外，与已开发的亲和材料相比，这种新型整体材料对抗体具有良好的抗体特异性选择性、相当或甚至更好的结合能力(对于干燥材料，最大静态结合能力约为119.3 mg/g，动态结合能力约为17.05 mg/g)，对胰酶消化的抵抗力可接受，对非特异性蛋白质的吸附可忽略不计。与相应的环肽琼脂糖凝胶材料相比，HT25环肽整体材料的洗脱条件较温和，可以有效地防止抗体的聚集和变性。将这种新型材料成功地应用于不同细胞培养液或人血清中的抗体抗体的亲和纯化。
The rapidly increasing applications of monoclonal antibodies (mAbs) in therapy have necessitated the development of mAb production and purification technologies for both academic and industrial usage. Herein, a histidine-tagged cyclic peptide (HHHHHHGSGSGSDC*AWHLGELVWC*T, the disulfide-bonded cysteines of which are indicated by asterisks, named HT25-cyclopeptide) functionalized monolithic material was developed by the metal ion chelation-based approach. The resulting material possessed suitable affinity and peptide ligand density (13.8 mg peptide ligand per mL of material), good porosity (67.1 %), acceptable specific surface area (52.95 m2/g), and lots of macropores (4.13 μm). Moreover, excellent antibody-specific selectivity, comparable or even better binding capacity (for dried material, maximum static binding capacity and dynamic binding capacity are about 119.3 mg/g and 17.05 mg/g, respectively) for antibody compared to previously developed affinity materials, acceptable resistance to trypsin digestion, and negligible nonspecific protein adsorption, were also achieved on this novel monolithic material. Compared with the corresponding cyclic peptide-based sepharose material, milder elution conditions were employed for the HT25-cyclopeptide-based monolithic material, which could effectively prevent the aggregation and denaturation of the enriched antibodies. This novel material was then successfully applied to the affinity enrichment and purification of mAbs (including infliximab and rituximab) in different cell culture media or IgG in human serum.