Gomes Cláudia Sg, Fashina Adedayo, Fernández-Castané Alfred, Overton Timothy W, Hobley Timothy J, Theodosiou Eirini, Thomas Owen Rt
Institute of Biotechnology and Biomedicine Technical University of Denmark Lyngby Denmark.
School of Chemical Engineering University of Birmingham Birmingham UK.
J Chem Technol Biotechnol. 2018 Jul;93(7):1901-1915. doi: 10.1002/jctb.5599. Epub 2018 Mar 30.
The extraction of biopharmaceuticals from plasma and serum often employs overly complicated antiquated procedures that can inflict serious damage on especially prone protein targets and which afford low purification power and overall yields. This paper describes systematic development of a high-gradient magnetic fishing process for recovery of immunoglobulins from unclarified antiserum.
Non-porous superparamagnetic particles were transformed into hydrophobic-charge induction adsorbents and then used to recover immunoglobulins from rabbit antiserum feedstocks. Comprehensive characterisation tests conducted with variously diluted clarified antiserum on a magnetic rack revealed that immunoglobulin binding was rapid (equilibrium reached in <45 s), strong (K < 0.1 mg mL), of high capacity (Q = 214 mg g), and pH and ionic strength dependent. In a high-gradient magnetic fishing process conducted with the same adsorbent, and a conventional 'magnetic filter + recycle loop' arrangement, >72% of the immunoglobulin present in an unclarified antiserum feed was recovered in 0.5 h in >3-fold purified form.
Fast magnetic particle based capture of antibodies from an unclarified high-titre feed has been demonstrated. Efficient product recovery from ultra-high titre bioprocess liquors by high-gradient magnetic fishing requires that improved magnetic adsorbents displaying high selectivity, ultra-high capacity and operational robustness are used with 'state-of-the-art' rotor-stator magnetic separators. © 2018 The Authors. published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.
从血浆和血清中提取生物制药产品通常采用过于复杂的陈旧程序,这些程序可能会对特别容易受损的蛋白质靶点造成严重损害,而且纯化能力和总收率较低。本文描述了一种用于从未澄清抗血清中回收免疫球蛋白的高梯度磁捕法的系统开发。
将无孔超顺磁性颗粒转化为疏水电荷诱导吸附剂,然后用于从兔抗血清原料中回收免疫球蛋白。在磁架上对不同稀释度的澄清抗血清进行的综合表征测试表明,免疫球蛋白结合迅速(在<45秒内达到平衡)、牢固(K<0.1mg/mL)、容量高(Q=214mg/g),且依赖于pH值和离子强度。在使用相同吸附剂和传统“磁滤器+循环回路”装置进行的高梯度磁捕过程中,未澄清抗血清原料中72%以上的免疫球蛋白在0.5小时内以超过3倍的纯化形式被回收。
已证明基于磁性颗粒从未澄清的高滴度原料中快速捕获抗体。通过高梯度磁捕从未澄清的超高滴度生物工艺液中高效回收产品,需要使用具有高选择性、超高容量和操作稳健性的改进型磁性吸附剂以及“先进的”转子-定子磁选机。©2018作者。由John Wiley & Sons Ltd代表化学工业协会出版。