多克隆抗体
化学
色谱法
胶束
肺表面活性物质
溶解
单克隆抗体
产量(工程)
动态光散射
单体
抗体
纳米颗粒
生物化学
材料科学
有机化学
生物
纳米技术
聚合物
水溶液
冶金
免疫学
作者
Gunasekaran Dhandapani,Ellen Wachtel,Guy Patchornik
出处
期刊:Nano select
[Wiley]
日期:2023-03-27
卷期号:4 (6): 386-394
被引量:1
标识
DOI:10.1002/nano.202200251
摘要
Abstract As downstream purification and separation technologies progress towards raising the concentration of therapeutic‐grade monoclonal antibodies (mAbs) in cell culture, downstream processing has begun to face increased difficulty in efficiently coping with such high immunoglobulin G (IgG) titers (≤25 mg mL −1 ). In the current study, we demonstrate the ability of a non‐chromatographic, ligand‐free procedure to recover almost quantitatively (84–99% yield, by densitometry) polyclonal, human IgG present at high concentrations (15–25 mg mL −1 ) in E. coli lysate. Instead of chromatographic media and columns, we use conjugated, mixed‐micelles comprising non‐ionic detergents, tyrosine monomers, and the amphiphilic [(bathophenanthroline) 3 :Fe 2+ ] complex. Capture and extraction processes are performed at pH 6.5–7, thereby avoiding antibody exposure to acidic, potentially denaturing conditions. Recovered IgG is monomeric as determined by dynamic light scattering (DLS). Process upscaling from 0.1 to 5 mL requires only proportional increase in all reagents and does not affect overall yield or antibody purity.
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