Reversible interference of Fe3+ with monoclonal antibody analysis in cation exchange columns

Rao, Srinivasa; Pohl, Christopher A.

doi:10.1016/j.ab.2010.10.026

Cited by 14 publications

(3 citation statements)

References 17 publications

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“…Furthermore, polyprotic citrate in the biologic pH range carries a full negative charge (similar to phosphate). It has been discussed that the full negative charges on citrate and phosphate prevents effective buffering interaction with the deprotonated CEX phase, and there is also the possibility that citrate can intercalate the protein, changing its conformation and fundamentally affecting IEX separation . We have not found that polyvalent salts have poor performance as chromatography buffer nor did we have an issue with microbial growth; however, since our purpose was to develop a shorter PAT method that would be connected to the continuous process for long durations, we considered alternatives to citrate.…”

Section: Results and Discussionmentioning

confidence: 99%

On-Line Ion Exchange Liquid Chromatography as a Process Analytical Technology for Monoclonal Antibody Characterization in Continuous Bioprocessing

et al. 2017

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Combining process analytical technology (PAT) with continuous production provides a powerful tool to observe and control monoclonal antibody (mAb) fermentation and purification processes. This work demonstrates on-line liquid chromatography (on-line LC) as a PAT tool for monitoring a continuous biologics process and forced degradation studies. Specifically, this work focused on ion exchange chromatography (IEX), which is a critical separation technique to detect charge variants. Product-related impurities, including charge variants, that impact function are classified as critical quality attributes (CQAs). First, we confirmed no significant differences were observed in the charge heterogeneity profile of a mAb through both at-line and on-line sampling and that the on-line method has the ability to rapidly detect changes in protein quality over time. The robustness and versatility of the PAT methods were tested by sampling from two purification locations in a continuous mAb process. The PAT IEX methods used with on-line LC were a weak cation exchange (WCX) separation and a newly developed shorter strong cation exchange (SCX) assay. Both methods provided similar results with the distribution of percent acidic, main, and basic species remaining unchanged over a 2 week period. Second, a forced degradation study showed an increase in acidic species and a decrease in basic species when sampled on-line over 7 days. These applications further strengthen the use of on-line LC to monitor CQAs of a mAb continuously with various PAT IEX analytical methods. Implementation of on-line IEX will enable faster decision making during process development and could potentially be applied to control in biomanufacturing.

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Section: Results and Discussionmentioning

confidence: 99%

On-Line Ion Exchange Liquid Chromatography as a Process Analytical Technology for Monoclonal Antibody Characterization in Continuous Bioprocessing

et al. 2017

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“…When running SEC methods in the first dimension of 2D-LC setups with mobile phases containing a high amount of chloride anions, regular stainless steel systems will be corroded over time and release metals in the sample flow path. In the meantime, the release of iron has been shown to induce mAb adsorption on CEX columns (74). In addition, non-specific interactions between the stainless steel parts of LC systems and phosphate-containing buffers are well-known, but may occur with proteins too (75).…”

Section: Two-dimensional Liquid Chromatography/mass Spectrometry (2d-mentioning

confidence: 99%

Multi-dimensional LC-MS: the next generation characterization of antibody-based therapeutics by unified online bottom-up, middle-up and intact approaches

Camperi

Goyon

Guillarme

et al. 2021

Analyst

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“…Metal contamination may be reversed by flushing with chelating agents such as oxalic acid dihydrate (Rao & Pohl, 2011). Also, stainless steel systems may require periodic passivation for reliable usage (Collins et al, 2000b).…”

Section: Metal Contaminationmentioning

confidence: 99%

Innovations in biotechnology

Gartland

Beccari

Bruschi

et al. 2015

Journal of Biotechnology

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Reversible interference of Fe3+ with monoclonal antibody analysis in cation exchange columns

Cited by 14 publications

References 17 publications

On-Line Ion Exchange Liquid Chromatography as a Process Analytical Technology for Monoclonal Antibody Characterization in Continuous Bioprocessing

On-Line Ion Exchange Liquid Chromatography as a Process Analytical Technology for Monoclonal Antibody Characterization in Continuous Bioprocessing

Multi-dimensional LC-MS: the next generation characterization of antibody-based therapeutics by unified online bottom-up, middle-up and intact approaches

Innovations in biotechnology

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