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Characterization of human papillomavirus 6b L1 virus-like particles isolated from silkworms using capillary zone electrophoresis.

Characterization of human papillomavirus 6b L1 virus-like particles isolated from silkworms using capillary zone electrophoresis.

Journal of bioscience and bioengineering (2014-04-04)

Thomas Hahne, Muthukutty Palaniyandi, Tatsuya Kato, Peter Fleischmann, Hermann Wätzig, Enoch Y Park

ABSTRACT

Human papillomavirus 6b L1 virus-like particles (VLPs) were successfully expressed using Bombyx mori nucleopolyhedrovirus (BmNPV) bacmid expression system and rapidly purified using size exclusion chromatography after ultracentrifugation procedure and characterized by capillary zone electrophoresis (CZE). The average capillary electrophoresis migration time was 11 min with the relative standard deviation (RSD) of 0.3% of human papillomavirus 6b L1 VLPs. After this threefold fractionation, the CZE samples were still further investigated by dynamic light scattering and immuno blotting. The versatile technique, CZE not only proved to be a valuable tool for VLP characterization, but was also found to be reliable and precise. Thus CZE will also be an important option for the quality control of VLPs in pharmaceutical research level.

MATERIALS

Product Number

Brand

Product Description

Supelco

Sodium hydroxide concentrate, 0.1 M NaOH in water (0.1N), Eluent concentrate for IC

Sigma-Aldrich

Sodium chloride solution, 0.1 M

Sigma-Aldrich

Sodium hydroxide solution, 7 M

Sigma-Aldrich

Sodium chloride solution, 1 M

Sigma-Aldrich

Sodium hydroxide solution, 6 M

Sigma-Aldrich

Potassium chloride solution, 0.01 M

Sigma-Aldrich

Sodium hydroxide solution, 0.1 M

Sigma-Aldrich

Sodium hydroxide solution, 1 M

Sigma-Aldrich

Sodium hydroxide, JIS special grade, ≥96.0%

Sigma-Aldrich

Sodium hydroxide solution, 4 M

Sigma-Aldrich

Sodium chloride, SAJ first grade, ≥99.0%

Sigma-Aldrich

Sodium chloride, JIS special grade, ≥99.5%

Sigma-Aldrich

Sodium phosphate dibasic solution, BioUltra, 0.5 M in H₂O

Supelco

Phosphate Standard for IC, TraceCERT^®, 1000 mg/L phosphate in water (nominal concentration)

SAFC

Sodium chloride solution, 5 M

Sigma-Aldrich

Sodium chloride, BioPerformance Certified, ≥99% (titration), suitable for insect cell culture, suitable for plant cell culture

Sigma-Aldrich

Potassium chloride, BioUltra, for molecular biology, ≥99.5% (AT)

Sigma-Aldrich

Acetanilide, puriss. p.a., ≥99.5% (CHN)

Sigma-Aldrich

Potassium chloride, tested according to Ph. Eur.

Sigma-Aldrich

Sodium hydroxide solution, BioUltra, for molecular biology, 10 M in H₂O

Supelco

Sodium hydroxide solution, 49-51% in water, eluent for IC

Sigma-Aldrich

Sodium hydroxide, BioUltra, for luminescence, ≥98.0% (T), pellets

Sigma-Aldrich

Acetanilide, purified by sublimation, ≥99.9%

Sigma-Aldrich

Sodium chloride, random crystals, optical grade, 99.9% trace metals basis

Sigma-Aldrich

Sodium chloride, 99.999% trace metals basis

Sigma-Aldrich

Acetanilide, 99%

Sigma-Aldrich

Potassium chloride, AnhydroBeads^™, −10 mesh, 99.99% trace metals basis

Sigma-Aldrich

Sodium chloride, AnhydroBeads^™, −10 mesh, 99.999% trace metals basis

Sigma-Aldrich

Potassium chloride, AnhydroBeads^™, −10 mesh, 99.999% trace metals basis

Sigma-Aldrich

3-Ethyl-2,4-pentanedione, mixture of tautomers, 98%