Format

Send to

Choose Destination
Microbiology. 1996 Jan;142 ( Pt 1):57-62.

Purification of meningococcal lipo-oligosaccharide by FPLC techniques.

Author information

1
Division of Bacteriology, National Institute for Biological Standards and Control, South Mimms, Potters Bar, Herts, UK.

Abstract

A rapid and efficient method for the preparation of highly pure meningococcal lipo-oligosaccharide (LOS) was developed. This used a Superose 6 column on a FPLC system to purify LOS from phenol-water extracts of cell lysates of Neisseria meningitidis. The purest LOS preparations, with no detectable protein contamination and less than 0.5% (w/w) residual RNA, were obtained when cell lysates had been treated with RNase ONE before phenol extraction and chromatographic separation. Preparations that had received no ribonuclease treatment had 2-3% residual RNA contamination and predigestion of samples with RNase A, which only partially degraded the RNA present in the crude extracts, resulted in LOS samples contaminated with 15-20% residual RNA. The LOS purified from RNase ONE-treated extracts was highly endotoxic, and showed no reduction in antibody binding or specific endotoxin activity compared to unpurified material. Approximately 80% of the LOS applied to the chromatography column was recovered as purified material.

PMID:
8581170
DOI:
10.1099/13500872-142-1-57
[Indexed for MEDLINE]

Supplemental Content

Full text links

Icon for Ingenta plc
Loading ...
Support Center