Biosynthetic radiolabeling of bacterial lipopolysaccharide to high specific activity

Robert S. Munford; Linda C. DeVeaux; J. E. Cronan; Paul D. Rick

doi:10.1016/0022-1759(92)90164-O

Biosynthetic radiolabeling of bacterial lipopolysaccharide to high specific activity

Robert S. Munford, Linda C. DeVeaux, J. E. Cronan, Paul D. Rick

Research output: Contribution to journal › Article › peer-review

Abstract

We describe a method for producing radiolabeled lipopolysaccharide (LPS) by incorporating [³H]acetate into an aceEF, gltA strain of Escherichia coli K12. The LPS has substantially greater specific radioactivity (2 μCi per μg LPS, or approximately 8 Ci/mmol) than has been reported previously for biosynthetically radiolabeled LPS. The ³H is incorporated into the fatty acyl chains of the lipid A moiety. LPS prepared by this method has several attractive features for biological studies, including native structure and bioactivity, long radioactive half-life, and high specific activity.

Original language	English (US)
Pages (from-to)	115-120
Number of pages	6
Journal	Journal of Immunological Methods
Volume	148
Issue number	1-2
DOIs	https://doi.org/10.1016/0022-1759(92)90164-O
State	Published - 1992

Keywords

Endotoxin
Lipid A
Lipopolysaccharide
Radiolabel

ASJC Scopus subject areas

Immunology and Allergy
Immunology

Online availability

10.1016/0022-1759(92)90164-O

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title = "Biosynthetic radiolabeling of bacterial lipopolysaccharide to high specific activity",

abstract = "We describe a method for producing radiolabeled lipopolysaccharide (LPS) by incorporating [3H]acetate into an aceEF, gltA strain of Escherichia coli K12. The LPS has substantially greater specific radioactivity (2 μCi per μg LPS, or approximately 8 Ci/mmol) than has been reported previously for biosynthetically radiolabeled LPS. The 3H is incorporated into the fatty acyl chains of the lipid A moiety. LPS prepared by this method has several attractive features for biological studies, including native structure and bioactivity, long radioactive half-life, and high specific activity.",

keywords = "Endotoxin, Lipid A, Lipopolysaccharide, Radiolabel",

author = "Munford, {Robert S.} and DeVeaux, {Linda C.} and Cronan, {J. E.} and Rick, {Paul D.}",

note = "Funding Information: This work was supported by US Public Health Service grant AI18188 from the National Institute of Allergy and Infectious Diseases.",

year = "1992",

doi = "10.1016/0022-1759(92)90164-O",

language = "English (US)",

volume = "148",

pages = "115--120",

journal = "Journal of Immunological Methods",

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T1 - Biosynthetic radiolabeling of bacterial lipopolysaccharide to high specific activity

AU - Munford, Robert S.

AU - DeVeaux, Linda C.

AU - Cronan, J. E.

AU - Rick, Paul D.

N1 - Funding Information: This work was supported by US Public Health Service grant AI18188 from the National Institute of Allergy and Infectious Diseases.

PY - 1992

Y1 - 1992

N2 - We describe a method for producing radiolabeled lipopolysaccharide (LPS) by incorporating [3H]acetate into an aceEF, gltA strain of Escherichia coli K12. The LPS has substantially greater specific radioactivity (2 μCi per μg LPS, or approximately 8 Ci/mmol) than has been reported previously for biosynthetically radiolabeled LPS. The 3H is incorporated into the fatty acyl chains of the lipid A moiety. LPS prepared by this method has several attractive features for biological studies, including native structure and bioactivity, long radioactive half-life, and high specific activity.

AB - We describe a method for producing radiolabeled lipopolysaccharide (LPS) by incorporating [3H]acetate into an aceEF, gltA strain of Escherichia coli K12. The LPS has substantially greater specific radioactivity (2 μCi per μg LPS, or approximately 8 Ci/mmol) than has been reported previously for biosynthetically radiolabeled LPS. The 3H is incorporated into the fatty acyl chains of the lipid A moiety. LPS prepared by this method has several attractive features for biological studies, including native structure and bioactivity, long radioactive half-life, and high specific activity.

KW - Endotoxin

KW - Lipid A

KW - Lipopolysaccharide

KW - Radiolabel

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JO - Journal of Immunological Methods

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Biosynthetic radiolabeling of bacterial lipopolysaccharide to high specific activity

Abstract

Keywords

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