Corrigendum to “Polishing monoclonal antibody using pH-responsive TiO2/polysulfone membrane in dual size-exclusion strategy” [Sep. Purif. Technol. 213 (2019) 359–356]

Background:: Protein A affinity chromatography is often employed as the most crucial purification step for monoclonal antibodies to achieve high yield with purity and throughput requirements. Introduction:: Protein A, also known as Staphylococcal protein A (SPA) is found in the cell wall of the bacteria staphylococcus aureus. It is one of the first discovered immunoglobulin binding molecules and has been extensively studied since the past few decades. The efficiency of Protein A affinity chromatography to purify a recombinant monoclonal antibody in a cell culture sample has been evaluated, which removes 99.0% of feed stream impurities. Materials and Method:: We have systematically evaluated the purification performance by using a battery of analytical methods SDS-PAGE (non-reduced and reduced sample), Cation Exchange Chromatography (CEX), Size-exclusion chromatography (SEC), and Reversed phased-Reduced Chromatography for a CHO-derived monoclonal antibody. Results and Discussion:: The analytical test was conducted to determine the impurity parameter, Host Cell Contaminating Proteins (HCP). It was evaluated to be 0.015ng/ml after the purification step; while initially, it was found to be 24.431ng/ml. Conclusion:: The tests showed a distinct decrease in the level of different impurities after the chromatography step. It can be concluded that Protein A chromatography is an efficient step in the purification of monoclonal antibodies.

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Development of capillary size exclusion chromatography for the analysis of monoclonal antibody fragments extracted from human vitreous humor

Journal of Chromatography A ◽

10.1016/j.chroma.2012.10.051 ◽

2012 ◽

Vol 1270 ◽

pp. 111-117 ◽

Cited By ~ 7

Author(s):

Jennifer C. Rea ◽

Yun Lou ◽

Joel Cuzzi ◽

Yuhua Hu ◽

Isabella de Jong ◽

...

Keyword(s):

Monoclonal Antibody ◽

Size Exclusion Chromatography ◽

Vitreous Humor ◽

Antibody Fragments ◽

Size Exclusion ◽

Exclusion Chromatography ◽

Capillary Size

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Comparison of Three Complementary Analytical Techniques for the Evaluation of the Biosimilar Comparability of a Monoclonal Antibody and an Fc-Fusion Protein

Frontiers in Chemistry ◽

10.3389/fchem.2021.782099 ◽

2021 ◽

Vol 9 ◽

Author(s):

Alice Demelenne ◽

Arij Ben Yahia ◽

Delphine Lempereur ◽

Jacques Crommen ◽

Anne-Catherine Servais ◽

...

Keyword(s):

Monoclonal Antibody ◽

Fusion Protein ◽

Method Development ◽

Reference Product ◽

Degradation Products ◽

Stress Conditions ◽

Size Exclusion ◽

Mild Stress ◽

Molecular Weight Determination ◽

Fc Fusion Protein

In this work, a monoclonal antibody, adalimumab, and an Fc-fusion protein, etanercept, were studied and compared to one of their biosimilars. Samples submitted to stress conditions (agitation and high temperature) were used for method development. The developed methods were also applied to samples reduced by beta-mercaptoethanol to evaluate their capability to distinguish the expected species. Capillary gel electrophoresis (CGE), reversed-phase liquid chromatography (RPLC), and size-exclusion chromatography (SEC) methods coupled with UV detection were used to analyze the biopharmaceuticals. Their complementarity was investigated. For further molecular weight determination, SEC-multi angle light scattering and RPLC-quadrupole time-of-flight were occasionally used. For adalimumab, a larger amount of fragments and aggregates was observed in the biosimilar compared with the reference product. For etanercept, more related species were found in the reference product. Those three separation techniques showed good complementarity. Indeed, RPLC enabled the separation of hydrophilic and hydrophobic degradation products. CGE provided good selectivity for several adalimumab fragments, and SEC was useful for the analysis of aggregates and certain fragments that cannot be separated by the other approaches. Moreover, those formulations were submitted to mild stress conditions (30°C, 300 rpm for 4 h) that mimic shipping conditions. No additional peak was found under these conditions for the two studied biopharmaceuticals.

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Characterization of a monoclonal antibody directed against the biologically active site of human interleukin 1.

Journal of Experimental Medicine ◽

10.1084/jem.163.2.463 ◽

1986 ◽

Vol 163 (2) ◽

pp. 463-468 ◽

Cited By ~ 13

Author(s):

A Köck ◽

M Danner ◽

B M Stadler ◽

T A Luger

Keyword(s):

Monoclonal Antibody ◽

Active Site ◽

Biological Effects ◽

Interleukin 1 ◽

Biologically Active ◽

Size Exclusion ◽

Fibroblast Proliferation ◽

Initial Activity ◽

Exclusion Chromatography

Human IL-1 was successfully used to produce an anti-IL-1 mAb. Anti-IL-1 (IgG2a) blocked IL-1-mediated thymocyte and fibroblast proliferation, but did not interfere with the biological effects of other lymphokines, such as IL-2 or IL-3. The antibody immunoprecipitated biosynthetically radiolabeled 33, 17, and 4 kD IL-1. An immunoadsorbent column yielded 20% of initial activity, and upon HPLC size-exclusion chromatography, affinity-purified IL-1 had a molecular mass of approximately 4 kD. These results provide first evidence of a monoclonal anti-IL-1 that reacts with different species of IL-1 and apparently binds to an epitope close to the active site of IL-1. Thus, anti-IL-1 IgG may be very helpful for further investigations of the molecular as well as biological characteristics of IL-1 and related mediators.

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