IMPROVED ION-EXCHANGED HPLC METHOD IN mAb USING pH GRADIENT AND ITS COMPARISON WITH cIEF

Wang, Feng; Peklansky, Brian; Anderson, Carrie L.; Wang, Yang; Rustandi, Richard R.

doi:10.1080/10826076.2011.619035

Cited by 12 publications

(6 citation statements)

References 17 publications

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“…Mobile phase A consisted of 10 mM sodium phosphate, pH 7.2 and mobile phase B consisting of 10 mM sodium phosphate with 0.5 M sodium chloride, pH 7.2. Samples were diluted to 1 mg/mL in mobile phase A and were injected onto the column and detected using fluorescence with an excitation wavelength at 280 nm and an emission wavelength at 350 nm . The dengue samples were separated using a gradient of 0–60% mobile phase B in 20 min at 1 mL/min flow rate.…”

Section: Methodsmentioning

confidence: 99%

Development of a capillary zone electrophoresis method for dose determination in a tetravalent dengue vaccine candidate

2015

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Dengue is known to cause morbidity and mortality worldwide and currently there is neither available specific therapeutics to treat nor a vaccine to prevent this disease. Although efforts are being made, development of a vaccine against this disease remains challenging. Hawaii Biotech Inc developed a recombinant subunit envelope protein-based vaccine against all four serotypes produced in Drosophila S2 cells which were transferred over to Merck in 2010. Each subunit of the four dengue serotypes contains the N-terminal 80% of the amino acids comprising the envelope protein (DEN-80E). A Phase 1 study using only monovalent DEN1-80E was done by Hawaii Biotech Inc and most recently, a Phase 1 clinical trial of the tetravalent DEN-80E formulation (V180) was conducted. Here, we report the development of a dose assay for the tetravalent dengue vaccine-containing subunit protein of DEN1-80E, DEN2-80E, DEN3-80E, and DEN4-80E using various separation methods such as HPLC and CE. Based on the results of the comparison, the CZE separation was chosen as the most suitable method to perform the dose assay for the tetravalent dengue vaccine.

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Section: Methodsmentioning

confidence: 99%

Development of a capillary zone electrophoresis method for dose determination in a tetravalent dengue vaccine candidate

2015

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“…For example, ProPac ® WCX-10 column (Dionex) is a weak cation exchange column commonly used for the separation and analysis of mAbs, in which positively charged ions on exposed protein surface bind to a negatively charged functional group on column and are eluted by a salt or pH gradient [13][14]. Consequently, the method is more sensitive to the protein structure, i.e., the extent of exposure and arrangement of the charge groups on proteins.…”

Section: Ion Exchange Chromatography (Iec)mentioning

confidence: 99%

7.2 Analytical Techniques and Quality Control of Protein Therapeutics

Que¹,

Wang²,

Kan³

2014

Animal Cell Biotechnology

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“…Wang et al. investigated mAb charge variants applying salt gradient CEX and icIEF. A higher resolution could be achieved with icIEF.…”

Section: Introductionmentioning

confidence: 99%

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

Kahle

Wätzig

2018

Electrophoresis

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Biopharmaceuticals are increasingly present in pharmaceutical therapy. These highly complex molecules are still challenging for development and quality control. Charge heterogeneity is next to size heterogeneity, glycan analysis, or peptide mapping one main aspect of their thorough characterization. This review focuses on the charge-based analysis of biopharmaceuticals through CE. An overview is given about the three separation techniques cIEF, imaged cIEF and CZE for the analyses of charge variants, emphasizing assays and their precision. Especially the developments in the experimental conditions have been given a high priority: sample preparation, capillary properties, anolytes and catholytes, carrier ampholytes, sacrificing agents, solubility enhancement, isoelectric point marker, mobilization techniques, and detection modes have been discussed. CIEF shows a particularly high heterogeneity in its methods. Therefore, a design space is suggested that states the most important adjustable parameters and their range. This enables an adequate design of experiments for individual sample separations, in order to accelerate and simplify method developments.

show abstract

IMPROVED ION-EXCHANGED HPLC METHOD IN mAb USING pH GRADIENT AND ITS COMPARISON WITH cIEF

Cited by 12 publications

References 17 publications

Development of a capillary zone electrophoresis method for dose determination in a tetravalent dengue vaccine candidate

Development of a capillary zone electrophoresis method for dose determination in a tetravalent dengue vaccine candidate

7.2 Analytical Techniques and Quality Control of Protein Therapeutics

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

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