Journal article
Method development and validation of capillary sodium dodecyl sulfate gel electrophoresis for the characterization of a monoclonal antibody
Journal of pharmaceutical and biomedical analysis, v 53(5), pp 1236-1243
2010
PMID: 20719453
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
A capillary sodium dodecyl sulfate gel electrophoresis (cSDS) method has been developed and qualified for purity and impurity analysis of monoclonal antibodies. This method was optimized and qualified for the analysis of monoclonal antibody (mAb1) under reduced and non-reduced conditions.
Some of the sample preparation parameters including sample buffer pH, incubation temperature and duration, alkylation conditions with iodoacetamide (IAM), and reduction conditions with 2-mercaptoethanol (2-ME) were optimized. It was observed that under slightly acidic conditions (pH 5.5–6.5) the thermally induced fragmentation of non-reduced mAb1 was greatly decreased. As such, a citrate–phosphate buffer at pH 6.5 was used for sample preparation to replace the original Beckman sample buffer (pH 9.0). The optimal sample preparation conditions were found to be as follows: (1) incubation temperature and duration (reduced and non-reduced), 65
°C for 5
min; (2) alkylation condition, 10
μL of 0.25
M IAM; (3) reduction condition, 10
μL of 5-fold diluted 2-ME.
The method was qualified by evaluating specificity, accuracy, precision, limit of quantitation (LOQ), and linearity. The method exhibited no interference from sample buffer matrix. The method was found to be linear, accurate, and precise in the range of 0.25–3.0
mg/mL protein concentration. The LOQ of the method was determined to be 0.02
mg/mL for reduced and non-reduced mAb1. In addition, some aspects of sample stability were examined during qualification.
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Details
- Title
- Method development and validation of capillary sodium dodecyl sulfate gel electrophoresis for the characterization of a monoclonal antibody
- Creators
- Junge Zhang - Analytical Development, Johnson and Johnson Pharmaceutical Research and Development, Malvern, PA, USASudhir Burman - Analytical Development, Johnson and Johnson Pharmaceutical Research and Development, Malvern, PA, USASri Gunturi - Analytical Development, Johnson and Johnson Pharmaceutical Research and Development, Malvern, PA, USAJoe P Foley - Department of Chemistry, Drexel University, 3141 Chestnut Street, Philadelphia, PA 19104, USA
- Publication Details
- Journal of pharmaceutical and biomedical analysis, v 53(5), pp 1236-1243
- Publisher
- Elsevier
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Chemistry
- Web of Science ID
- WOS:000281576500015
- Scopus ID
- 2-s2.0-77955924947
- Other Identifier
- 991014877784404721
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- Collaboration types
- Industry collaboration
- Domestic collaboration
- Web of Science research areas
- Chemistry, Analytical
- Pharmacology & Pharmacy