Qualification of NISTmAb charge heterogeneity control assays

Abstract

The NISTmAb is a monoclonal antibody Reference Material from the National Institute of Standards and Technology; it is a class-representative IgG1κ intended serve as a pre-competitive platform for harmonization and technology development in the biopharmaceutical industry. The publication series of which this paper is a part describes NIST’s overall control strategy to ensure NISTmAb quality and availability over its lifecycle. In this paper, the development and qualification of methods for monitoring NISTmAb charge heterogeneity are described. Capillary zone electrophoresis (CZE) and capillary isoelectric focusing (CIEF) assays were optimized and evaluated as candidate assays for NISTmAb quality control. CIEF was found to be suitable as a structural characterization assay yielding information on the apparent pI of the NISTmAb. CZE was found to be better suited for routine monitoring of NISTmAb charge heterogeneity and was qualified for this purpose. This paper is intended to provide relevant details of NIST’s charge heterogeneity control strategy to facilitate implementation of the NISTmAb as a test molecule in the end user’s laboratory. Graphical AbstractRepresentative capillary zone electropherogram of the NIST monoclonal antibody (NISTmAb). The NISTmAb is a publicly available research tool intended to facilitate advancement of biopharmaceutical analytics. Electronic supplementary materialThe online version of this article (10.1007/s00216-017-0816-6) contains supplementary material, which is available to authorized users.

High‐resolution separation of monoclonal antibodies mixtures and their charge variants by an alternative and generic CZE method

“…The acidic variants were systematically higher in CIEF, but the generic method might overestimated the amount of acidic variants as discussed in . The choice of the data analysis software and integration parameters can also explain the differences in some cases but unlikely those observed in the present CZE study versus as the same operator manually integrated the peaks.…”

“…The acidic variants were systematically higher in CIEF, but the generic method might overestimated the amount of acidic variants as discussed in [23]. The choice of the data analysis software and integration parameters can also explain the differences in some cases [24] but unlikely those observed in the present CZE study versus [23] as the same operator manually integrated the peaks. The developed CZE method was then compared to the well-established CZE approach from He et al [16] involving the use of EACA, TETA and HPMC at pH = 5.7, with 17 representative FDA-and EMA-approved mAbs ( Fig.…”

“…Indeed, detrimental effects were observed by He et al at pHs higher than 5.2 with a relatively low basic mAb (pI = 7.3) [16]. Similarly, HPMC was replaced by hydroxypropyl cellulose (HPC) to improve the separation of basic antibodies charge variants [15] and a nonionic surfactant (polysorbate 20) to avoid a significant loss of resolution occurring after few injections [24].…”

Determination of protein charge variants with (imaged) capillary isoelectric focusing and capillary zone electrophoresis

“…In addition, a third method was calculated for the determination of a more acidic mAb, called “Mab2 specific compromise method.” This shows, that despite of the other two methods, a further optimization can be useful in specific cases. Turner and Schiel optimized the CZE method based on He et al. .…”

Application of affinity capillary electrophoresis for charge heterogeneity profiling of biopharmaceuticals

“…For this purpose, a CZE–CZE–MS approach was published recently . Due to the individual properties of different biopharmaceuticals, various modifications of the initial method from He and colleagues have been published . Goyon et al.…”