Fractionated charge variants of biosimilars: A review of separation methods, structural and functional analysis

Yüce, Meral; Sert, Fatma; Torabfam, Milad; Parlar, Ayhan; Gürel, Büşra; Çakır, Nilüfer; Dağlikoca, Duygu Emine; Khan, Mansoor A.; Çapan, Yılmaz

doi:10.1016/j.aca.2020.12.064

Cited by 20 publications

(17 citation statements)

References 113 publications

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“…Clogging by aggregation results from a continuous deposition of particles. The aggregated layer grows as a result of competition between hydrodynamic, diffusive, and colloidal effects [104,105]. There are experimental and computational approaches to preventing clogging in microfluidic channels for both single-pored and porous materials [106].…”

Section: Clog Avoidance In Oocsmentioning

confidence: 99%

Design and Fabrication of Organ-on-Chips: Promises and Challenges

Tajeddin

Mustafaoğlu

2021

Micromachines

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The advent of the miniaturization approach has influenced the research trends in almost all disciplines. Bioengineering is one of the fields benefiting from the new possibilities of microfabrication techniques, especially in cell and tissue culture, disease modeling, and drug discovery. The limitations of existing 2D cell culture techniques, the high time and cost requirements, and the considerable failure rates have led to the idea of 3D cell culture environments capable of providing physiologically relevant tissue functions in vitro. Organ-on-chips are microfluidic devices used in this context as a potential alternative to in vivo animal testing to reduce the cost and time required for drug evaluation. This emerging technology contributes significantly to the development of various research areas, including, but not limited to, tissue engineering and drug discovery. However, it also brings many challenges. Further development of the technology requires interdisciplinary studies as some problems are associated with the materials and their manufacturing techniques. Therefore, in this paper, organ-on-chip technologies are presented, focusing on the design and fabrication requirements. Then, state-of-the-art materials and microfabrication techniques are described in detail to show their advantages and also their limitations. A comparison and identification of gaps for current use and further studies are therefore the subject of the final discussion.

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Section: Clog Avoidance In Oocsmentioning

confidence: 99%

Design and Fabrication of Organ-on-Chips: Promises and Challenges

Tajeddin

Mustafaoğlu

2021

Micromachines

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“…40) IEX can provide an informative chromatogram, including information about the charge variation, glycoforms, and structural alternations. [41][42][43][44] IEX is often used for protein analysis; however, there are a limited number of reports of its application to ADC analyses published in the literature. Recently, IEX coupled with MS under native conditions was applied for antibody characterization by several groups.…”

Section: Iex-qtof/msmentioning

confidence: 99%

Recent Advances in Drug–Antibody Ratio Determination of Antibody–Drug Conjugates

Matsuda

Mendelsohn

2021

Chem. Pharm. Bull.

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Antibody-drug conjugates (ADCs) are biopharmaceuticals produced by chemically linking small molecules (payloads) to antibodies that possess specific affinity for the target cell. The ADCs currently on the commercially market are the result of a stochastic conjugation of highly-potent payloads to multiple sites on the monoclonal antibody, resulting in a heterogeneous drug-antibody ratio (DAR) and drug distribution. The heterogeneity inherent to ADCs not produced site-specifically may not only be detrimental to the quality of the drug but also is less-desirable from the perspective of regulatory science. An ideal method or unified approach used to measure the DAR for ADCs, a critical aspect of their analysis and characterization, has not yet been established in the ADC field and remains an often-challenging issue for bioanalytical chemists. In this review we describe, compare, and evaluate the characteristics of various DAR determination methods for ADCs featuring recently reported technologies. The future landscape of bioconjugate DAR analysis is also discussed.

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“…They are categorized as acidic, basic, and the main (neutral) variants based on the mAb charge characteristics. Charge variations usually do not affect the biological function of mAbs [22], however, there are proven examples, where the potency and efficacy of mAbs is affected by the presence of different charge variants formed by amino or carboxyl‐terminal modifications, fragmentations, glycosylations, oxidations, or deamidations located on specific amino acid residues [23]. Charge variants analysis is, therefore, one of the parameters that must be determined during antibody production, primarily in recombinant antibodies for use in therapy.…”

Section: Introductionmentioning

confidence: 99%

“…Charge variants analysis is, therefore, one of the parameters that must be determined during antibody production, primarily in recombinant antibodies for use in therapy. Capillary isoelectric focusing (cIEF), 2D PAGE, 2D LC have been recently suggested for charge variant analyses [23], but these techniques are not appropriate for preparative applications. The rise of mAb biosimilars has renewed the interest in mAb charge variants composition and fractionation on preparative scale, because biosimilars are required to have charge variant composition identical to the innovator product [23,24].…”

Section: Introductionmentioning

confidence: 99%

“…Capillary isoelectric focusing (cIEF), 2D PAGE, 2D LC have been recently suggested for charge variant analyses [23], but these techniques are not appropriate for preparative applications. The rise of mAb biosimilars has renewed the interest in mAb charge variants composition and fractionation on preparative scale, because biosimilars are required to have charge variant composition identical to the innovator product [23,24]. A cation exchange chromatography (CEX) usually enables partial separation of mAbs charge variants in a salt or a pH gradient [25–27], but baseline separation is rarely achieved.…”

Section: Introductionmentioning

confidence: 99%

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Sample displacement chromatography of monoclonal antibody charge variants and aggregates

2021

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The rise of biosimilar monoclonal antibodies has renewed the interest in monoclonal antibody (mAb) charge variants composition and separation. The sample displacement chromatography (SDC) has the potential to overcome the low separation efficiency and productivity associated with bind‐elute separation of mAb charge variants. SDC in combination with weak cation exchanging macroporous monolithic chromatographic column was successfully implemented for a separation of charge variants and aggregates of monoclonal IgG under overloading conditions. The charge variants composition was at‐line monitored by a newly developed, simple and fast analytical method, based on weak cation exchange chromatography. It was proven that basic charge variants acted as displacers of IgG molecules with lower pI, when the loading was performed 1 to 1.5 pH unit below the pI of acidic charge variants. The efficiency of the SDC process is flow rate independent due to a convection‐based mass transfer on the macroporous monolith. The productivity of the process at optimal conditions is 35 mg of purified IgG fraction per milliliters of monolithic support with 75–80% recovery. As such, an SDC approach surpasses the standard bind‐elute separation in the productivity for a factor of 3, when performed on the same column. The applicability of the SDC approach was confirmed for porous particle‐based column as well, but with 1.5 lower productivity compared to the monoliths.

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Fractionated charge variants of biosimilars: A review of separation methods, structural and functional analysis

Cited by 20 publications

References 113 publications

Design and Fabrication of Organ-on-Chips: Promises and Challenges

Design and Fabrication of Organ-on-Chips: Promises and Challenges

Recent Advances in Drug–Antibody Ratio Determination of Antibody–Drug Conjugates

Sample displacement chromatography of monoclonal antibody charge variants and aggregates

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