Boi Cristiana
Dipartimento di Ingegneria Chimica, Mineraria e delle Tecnologie Ambientali, Università degli Studi di Bologna, Viale Risorgimento 2, 40136 Bologna, Italy.
J Chromatogr B Analyt Technol Biomed Life Sci. 2007 Mar 15;848(1):19-27. doi: 10.1016/j.jchromb.2006.08.044. Epub 2006 Sep 22.
Downstream purification processes for monoclonal antibody production typically involve multiple steps; some of them are conventionally performed by bead-based column chromatography. Affinity chromatography with Protein A is the most selective method for protein purification and is conventionally used for the initial capturing step to facilitate rapid volume reduction as well as separation of the antibody. However, conventional affinity chromatography has some limitations that are inherent with the method, it exhibits slow intraparticle diffusion and high pressure drop within the column. Membrane-based separation processes can be used in order to overcome these mass transfer limitations. The ligand is immobilized in the membrane pores and the convective flow brings the solute molecules very close to the ligand and hence minimizes the diffusional limitations associated with the beads. Nonetheless, the adoption of this technology has been slow because membrane chromatography has been limited by a lower binding capacity than that of conventional columns, even though the high flux advantages provided by membrane adsorbers would lead to higher productivity. This review considers the use of membrane adsorbers as an alternative technology for capture and polishing steps for the purification of monoclonal antibodies. Promising industrial applications as well as new trends in research will be addressed.
单克隆抗体生产的下游纯化过程通常涉及多个步骤;其中一些传统上是通过基于珠子的柱色谱法进行的。蛋白A亲和色谱法是蛋白质纯化最具选择性的方法,传统上用于初始捕获步骤,以促进快速体积减少以及抗体的分离。然而,传统的亲和色谱法有一些该方法固有的局限性,它在柱内表现出缓慢的颗粒内扩散和高压降。基于膜的分离过程可用于克服这些传质限制。配体固定在膜孔中,对流流动使溶质分子非常接近配体,从而最大限度地减少了与珠子相关的扩散限制。尽管如此,这项技术的采用一直很缓慢,因为膜色谱法的结合能力低于传统柱,尽管膜吸附器提供的高通量优势会带来更高的生产率。本文综述了膜吸附器作为单克隆抗体纯化捕获和精制步骤的替代技术的应用。将探讨有前景的工业应用以及新的研究趋势。