Purification of Anti-Interleukin-6 Monoclonal Antibody Using Precipitation and Immobilized Metal-Ion Affinity Chromatography
Purification of Anti-Interleukin-6 Monoclonal Antibody Using Precipitation and Immobilized Metal-Ion Affinity Chromatography
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DOI：
10.1260/0263-6174.33.2.191
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发表时间：
2015-02-01
影响因子：
2.9
通讯作者：
Pessoa, Adalberto
中科院分区：
文献类型：
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作者：
Antunes Pereira Bresolin, Iara Rocha;Lazzarotto Bresolin, Igor Tadeu;Pessoa, Adalberto
单克隆抗体（MAb）已广泛应用于免疫诊断和临床测试，其应用需要高纯度。因此，单克隆抗体的下游加工需要几个阶段的纯化。本研究的目的是通过沉淀和固定化金属离子亲和层析（IMAC）相结合的方法从细胞培养上清液中回收和纯化抗白细胞介素6单克隆抗体，旨在其随后在诊断试剂盒中的应用。实验结果表明，可以通过用 20-40% 饱和度的硫酸铵沉淀，然后用固定化 Zn(II) 吸附在 IMAC 上来纯化 MAb。获得的单克隆抗体纯度为100%，纯化因子为24.2，证明了该方法的可行性。
Monoclonal antibodies (MAbs) have been widely used in immunodiagnostic and clinical tests, whose applications require a high level of purity. As a result, several stages of purification are required in the downstream processing of MAbs. The objective of this study was to recover and purify an anti-interleukin-6 MAb from cell culture supernatant by a combined method involving precipitation and immobilized metal-ion affinity chromatography (IMAC), aiming its subsequent application in diagnostic kits. Experimental results suggest that it is possible to purify MAbs by precipitation with ammonium sulphate at 20-40% saturation, followed by adsorption on IMAC with immobilized Zn(II). The MAbs were obtained with a purity of 100% and a purification factor of 24.2, proving the feasibility of the method.