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

RDD038

Sigma-Aldrich

Sodium phosphate dibasic, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%, free-flowing, Redi-Dri^™

71640

Sigma-Aldrich

Sodium phosphate dibasic, puriss. p.a., ACS reagent, anhydrous, ≥99.0% (T)

PHR1530

Supelco

Acetaminophen Related Compound D, Pharmaceutical Secondary Standard; Certified Reference Material

60220

Sigma-Aldrich

Potassium phosphate monobasic, buffer substance, anhydrous, puriss. p.a., ACS reagent, reag. ISO, reag. Ph. Eur., 99.5-100.5%

04243

Sigma-Aldrich

Potassium phosphate monobasic, meets analytical specification of Ph. Eur., NF, E340, anhydrous, 98-100.5% (calc. to the dried substance)

04276

Sigma-Aldrich

Sodium phosphate dibasic, puriss., meets analytical specification of Ph. Eur., BP, USP, FCC, E 339, anhydrous, 98-100.5% (calc. to the dried substance)

P0662

Sigma-Aldrich

Potassium phosphate monobasic, ACS reagent, ≥99.0%

S9763

Sigma-Aldrich

Sodium phosphate dibasic, ACS reagent, ≥99.0%

PHR1330

Supelco

Monobasic Potassium Phosphate, Pharmaceutical Secondary Standard; Certified Reference Material

S5136

Sigma-Aldrich

Sodium phosphate dibasic, BioReagent, suitable for cell culture, suitable for insect cell culture, ≥99.0%