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

43617

Supelco

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

38364

Supelco

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

S6191

Sigma-Aldrich

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

60126

Sigma-Aldrich

Potassium chloride, tested according to Ph. Eur.

60128

Sigma-Aldrich

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

71383

Sigma-Aldrich

Sodium chloride, tested according to Ph. Eur.

71376

Sigma-Aldrich

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

60136

Supelco

Potassium chloride solution, conductance standard B acc. to ISO 7888, 0.01 M KCl

60138

Supelco

Potassium chloride solution, conductance standard C acc. to ISO 7888, 0.001 M KCl

60142

Sigma-Aldrich

Potassium chloride solution, BioUltra, for molecular biology, ~1 M in H₂O

71386

Sigma-Aldrich

Sodium chloride solution, BioUltra, for molecular biology, ~5 M in H₂O

71687

Sigma-Aldrich

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

72068

Sigma-Aldrich

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

60134

Supelco

Potassium chloride solution, conductance standard A acc. to ISO 7888, 0.1 M KCl

60137

Supelco

Potassium chloride solution, for Ag/AgCl electrodes, ~3 M KCl, saturated with silver chloride

58221

Supelco

ISA (ionic strength adjustment solution: 1 M KCl), 1 M KCl

72064

Supelco

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

60135

Supelco

Potassium chloride solution, BioUltra, ~3 M in H₂O

71387

Supelco

Sodium chloride, reference material for titrimetry, certified by BAM, >99.5%

409316

Sigma-Aldrich

Potassium chloride, 99.999% trace metals basis

204439

Sigma-Aldrich

Sodium chloride, 99.999% trace metals basis

451029

Sigma-Aldrich

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

397229

Sigma-Aldrich

Acetanilide, purified by sublimation, ≥99.9%

397237

Sigma-Aldrich

Acetanilide, zone-refined, purified by sublimation, ≥99.95%

451940

Sigma-Aldrich

Sodium chloride-³⁵Cl, 99 atom % ³⁵Cl

378860

Sigma-Aldrich

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

S0817

Sigma-Aldrich

Sodium chloride solution, 0.85%

204099

Sigma-Aldrich

Potassium chloride, ≥99.99% trace metals basis

449989

Sigma-Aldrich

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

450006

Sigma-Aldrich

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