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Rapid preparative separation of monoclonal antibody charge variants using laterally-fed membrane chromatography
- Source :
- Journal of Chromatography B. 1073:27-33
- Publication Year :
- 2018
- Publisher :
- Elsevier BV, 2018.
-
Abstract
- Monoclonal antibodies undergo various forms of chemical transformation which have been shown to cause loss in efficacy and alteration in pharmacokinetic properties of these molecules. Such modified antibody molecules are known as variants. They also display physical properties such as charge that are different from intact antibody molecules. However, the difference in charge is very subtle and separation based on it is quite challenging. Charge variants are usually separated using ion-exchange column chromatography or isoelectric focusing. In this paper, we report a rapid and scalable method for fractionating monoclonal antibody charge variants, based on the use of cation exchange laterally-fed membrane chromatography (LFMC). Starting with a sample of monoclonal antibody hIgG1-CD4, three well-resolved fractions were obtained using either pH or salt gradient. These fractions were identified as acidic, neutral and basic variants. Each of these fractions contained intact heavy and light chains and so antibody fragmentation had no role in variant generation. The separation was comparable to that using column chromatography but was an order of magnitude faster.
- Subjects :
- 0301 basic medicine
medicine.drug_class
Clinical Biochemistry
Sodium Chloride
Immunoglobulin light chain
Monoclonal antibody
01 natural sciences
Biochemistry
Analytical Chemistry
03 medical and health sciences
Column chromatography
medicine
Humans
Molecule
Fragmentation (cell biology)
Chromatography
biology
Chemistry
Isoelectric focusing
010401 analytical chemistry
Antibodies, Monoclonal
Membranes, Artificial
Cell Biology
General Medicine
Chromatography, Ion Exchange
0104 chemical sciences
030104 developmental biology
Membrane
Immunoglobulin G
biology.protein
Antibody
Subjects
Details
- ISSN :
- 15700232
- Volume :
- 1073
- Database :
- OpenAIRE
- Journal :
- Journal of Chromatography B
- Accession number :
- edsair.doi.dedup.....39e8e33e0ca486d6eecac47e7a12a6af