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Updated: Sep 5, 2025
Capillary Electrophoresis Separation of Monoclonal Antibody Isoforms Using a Neutral Capillary
Published on: January 16, 2017
Collaborative study for the establishment of Ph. Eur. Human immunoglobulin for electrophoresis BRP batch 4
M-E Behr-Gross1, E Regourd1, W Holtkamp2
1European Directorate for the Quality of Medicines & HealthCare (EDQM), Department of Biological Standardisation, OMCL Network & HealthCare (DBO), Council of Europe, Strasbourg, France.
Insights
A new European Pharmacopoeia Biological Reference Preparation (BRP) for human immunoglobulin electrophoresis was established. This replacement batch ensures continued quality control for immunoglobulin analysis in pharmaceutical preparations.
Area of Science:
- Pharmacopoeia Standards
- Biological Reference Materials
- Protein Electrophoresis
Background:
- Diminished stocks of the European Pharmacopoeia (Ph. Eur.) Biological Reference Preparation (BRP) for Human immunoglobulin for electrophoresis necessitated a replacement.
- The European Directorate for the Quality of Medicines & HealthCare (EDQM) initiated an international collaborative study in 2020 under the Biological Standardisation Programme (BSP).
Purpose of the Study:
- To establish a replacement batch for the Human immunoglobulin for electrophoresis BRP.
- To verify the suitability of candidate reference preparations against Ph. Eur. monographs for human normal immunoglobulin.
Main Methods:
- An international collaborative study involving nineteen laboratories.
- Testing of candidate preparations using zone electrophoresis with cellulose acetate and/or agarose.
- Exploration of capillary zone electrophoresis (CZE) as a potential analytical method.
Main Results:
- The candidate preparation was found suitable for its intended purpose.
- The new batch was adopted as Human immunoglobulin for electrophoresis BRP batch 4.
- An assigned range for immunoglobulin content was established as 82.5% to 87.8% of total protein content.
Conclusions:
- The established Human immunoglobulin for electrophoresis BRP batch 4 meets the requirements for quality control.
- The study successfully ensured the continued availability of a critical reference standard for immunoglobulin analysis.
- Further investigation into CZE for immunoglobulin quantification may be warranted.
Abstract:
Due to the diminished stocks of the third batch of the European Pharmacopoeia (Ph. Eur.) Biological Reference Preparation (BRP) for Human immunoglobulin for electrophoresis, in 2020 the European Directorate for the Quality of Medicines & HealthCare (EDQM) initiated an international collaborative study for the establishment of a replacement batch. The study was run under the aegis of the Biological Standardisation Programme (BSP). Nineteen laboratories participated in the collaborative study to verify the suitability of the candidate reference preparations according to the Ph. Eur. monographs Human normal immunoglobulin for intravenous administration (0918), Human normal immunoglobulin for intramuscular administration (0338) and Human normal immunoglobulin for subcutaneous administration (2788) using the zone electrophoresis method with cellulose acetate and/or agarose as the testing medium. Capillary zone electrophoresis (CZE), a technique not yet included in monographs 0338, 0918 and 2788, was also used by some laboratories. The assignment of a value for immunoglobulin as a percentage of the total protein content could only be made for agarose electrophoresis and for CZE. The candidate preparation was found suitable for the intended purpose and was subsequently adopted by correspondence in May 2021 by the Ph. Eur. Commission as Human immunoglobulin for electrophoresis BRP batch 4 with an assigned range for immunoglobulin of 82.5 % to 87.8 % of the total protein content.
