宿主细胞蛋白质和抗体含有异构聚合物的多组件行为在蛋白质A染色学中
Soumitra Bhoyar1, Max Foster1, Abraham M Lenhoff1
1Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, DE, 19716, USA.
Journal of chromatography. A
|May 1, 2025
概括
蛋白质A染色学与去除杂质进行斗争,包括与单克隆抗体 (mAbs) 共同化的异质聚合物. 这项研究模拟了聚合行为,以改善mAbs.的净化策略.
科学领域:
- 生物制药制造业 生物制药制造业
- 染色学科学 染色学科学
- 蛋白质化学 蛋白质化学
背景情况:
- 蛋白A染色学对于单克隆抗体 (mAb) 净化至关重要,但与宿主细胞蛋白 (HCP) 和高分子量 (HMW) 物种等持久杂质作斗争.
- mAb-HCP异构聚合物是杂质共化的一个关键因素,需要改进的分离方法.
- 了解这些聚合物种在染色学过程中的行为对于提高净化效率至关重要.
研究的目的:
- 为了研究IgG1和IgG4抗体的持久聚合物种 (小和大) 的染色学行为.
- 开发和验证基于一般速率模型 (GRM) 的机械模型,用于预测蛋白A染色学期间的总和mAb行为.
- 提供对聚合清除策略和多元组件染色学建模的见解.
主要方法:
- 对小 (SA) 和大 (LA) 聚合物和mAbs的染色学行为的实验性确定,使用采集的细胞培养液 (HCCF) 和蛋白A化物.
- 开发一个多组件的一般速率模型 (GRM) 来描述色谱突破和化.
- 模型的验证使用实验色谱数据和共聚焦激光扫描显微镜用于粒子内吸收.
主要成果:
- 开发的GRM准确地预测了列出口和粒子内部吸收配置文件,与实验数据保持一致.
- 聚合物与mAb的结合亲和率被量化,显示了pH范围内的显著差异.
- 较大的聚合物与小聚合物和mAbs相比,具有较高的结合亲和率,特别是在较低的pH值下.
结论:
- 该研究成功模拟了mAb聚合物的染色学行为,为预测和优化净化过程提供了一个框架.
- 该模型表明,受pH影响的结合亲和力的差异是将聚合物从mAbs.分离出来的关键.
- 这些发现和开发的模型可以帮助提高mAb净化过程中的聚合物清除,并对染色学建模具有更广泛的意义.
更多相关视频
09:12A Protein Microarray Assay for Serological Determination of Antigen-specific Antibody Responses Following Clostridium difficile Infection
Published on: June 15, 2018
07:35Double Labeling Immunofluorescence using Antibodies from the Same Species to Study Host-Pathogen Interactions
Published on: July 10, 2021
相关概念视频
01:20Immunoprecipitation
5.2K
Immunoprecipitation, or IP, is a widely used technique that employs protein-antibody interactions to isolate proteins or protein complexes in their native state for studying protein-protein interactions, quaternary structures, or supramolecular complexes. Various modifications of the technique, including chromatin IP, cross-linking IP, and fluorescence IP, are commonly used.
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
Chromatin Immunoprecipitation
Chromatin immunoprecipitation, also known as ChIP, is used to study protein-DNA or...
5.2K
01:03Affinity Chromatography
469
Affinity chromatography is a powerful technique extensively utilized for separating and purifying specific biomolecules from complex mixtures. It capitalizes on the highly selective binding between an analyte and its counterpart, such as antibody-antigen interactions. The counterpart is immobilized on the stationary phase, forming an affinity column. The stationary phase typically consists of solid support, such as agarose or porous glass beads, immobilizing the affinity ligand. The mobile...
469
01:26Antibody Actions
1.0K
Antibodies, or immunoglobulins, are critical players in the immune system's arsenal against invading pathogens. Produced by B cells and plasma cells, their primary role is to detect and bind to specific antigens, molecules found on the surface of pathogens like bacteria or viruses. Beyond antigen recognition, antibodies perform several vital functions that contribute to immune defense.
Neutralization
Antibodies can bind to pathogens, preventing them from infecting host cells. This process...
Neutralization
Antibodies can bind to pathogens, preventing them from infecting host cells. This process...
1.0K