Functional monolithic platforms: Chromatographic tools for antibody purification

Abstract

Polymer monoliths are an efficient platform for antibody purification. The use of monoclonal antibodies (mAbs) and engineered antibody structures as therapeutics has increased exponentially over the past few decades. Several approaches use polymer monoliths to purify large quantities of antibody with defined clinical and performance requirements. Functional monolithic supports have attracted a great deal of attention as they offer practical advantages for antibody purification, such as more rapid analysis, smaller sample volume requirements and the opportunity for a greater target molecule enrichment. This review focuses on the development of synthetic and natural polymer-based monoliths for antibody purification. The materials and methods employed in monolith production are discussed, highlighting the properties of each system. We also review the structural characterization techniques available using monolithic systems and their performance under different chromatographic approaches to antibody capture and release. Finally, a summary of monolithic platforms developed for antibody separation is presented, as well as expected trends in research to solve current and future challenges in this field. This review comprises a comprehensive analysis of proposed solutions highlighting the remarkable potential of monolithic platforms.

Highly linear pH gradients for analyzing monoclonal antibody charge heterogeneity in the alkaline range: Validation of the method parameters

“…methionine oxidation, C-terminal lysine processing, deamidation of asparagine and isomerization of aspartic acid influence the behavior of mAb, as such they can be considered a critical quality attribute (CQA) [5,6]. The emergence of novel purification strategies demands sophisticated analytical methods to ensure comparability of the product [7][8][9][10][11], and such analytical methods should be validated for their intended purpose [12,13].…”

Technology advancements in antibody purification

“…In chromatographic terms, a monolith is defined as a porous, single-unit material incorporated into a chromatographic device. 72,73 Manufacturers and analytical laboratories have been interested in the use of monolithic supports owing to their associated low cost, high porosity, and ease of use. 72 Since its discovery, a number of different materials, including organic monoliths, silica monoliths, agarose monoliths, and cryogels have been tested for utilization in affinity monolith chromatography, and some of the materials have been tested for antibody purification.…”

“…72,73 Manufacturers and analytical laboratories have been interested in the use of monolithic supports owing to their associated low cost, high porosity, and ease of use. 72 Since its discovery, a number of different materials, including organic monoliths, silica monoliths, agarose monoliths, and cryogels have been tested for utilization in affinity monolith chromatography, and some of the materials have been tested for antibody purification. 74 Glycidyl methacrylate (GMA) and ethylene glycol dimethacrylate have been used for affinity (using ligands Protein-A, -L, and -G and poly-His) and ion-exchange purification of antibodies.…”

“…IgG, IgM, IgA, and antibody fragments have all been purified in varying purities using monolithic supports. 72, 75 Feng et al 76 used immobilized lectin (Concanavalin A) monolithic chromatography to successfully purify glycoproteins using an IMAC-based mechanism. A comprehensive review of monolithic supports used for antibody purification was recently published by Barroso et al 72 …”

Editorial: ESBES – European Society of Biochemical Engineering Sciences

“…The work of Gruber et al [4] is an interesting example of biocatalysis using native hosts (in this case Candida tenuis and Pichia stipites) as whole-cell catalysts, which significantly shortens process development time. In the area of biopharmaceutical technology we have several manuscripts on new separation technologies such as convective chromato graphy media for antibody purification [5] and stimuli-responsive magnetic nanoparticles for monoclonal antibody purification [6]. Bioreactor design is in particular important for new cellular products using stem cells.…”