TY - CHAP
T1 - Chromatographic analysis of the Escherichia coli polysialic acid capsule
AU - Steenbergen, Susan M.
AU - Vimr, Eric R.
PY - 2013
Y1 - 2013
N2 - Polysialic acid capsules are the major virulence factors in Escherichia coli K1, K92, and groups B and C meningococci. The sialic acid monomers (2-keto-3-deoxy-5-acetamido-7,8,9- D - glycero - D - galacto -nonulosonic acids) comprising these homopolymeric polysaccharide chains can be selectively modified with 1,2- diamino-4,5-methylenedioxy-benzene to produce highly fluorescent quinoxalinone derivatives distinguished by their elution times during reverse phase chromatography. Here, we describe methods to release the constituent capsular polysialic acid monomers, and to detect and quantify them by sensitive fluorometry. There are relatively few 2-keto acids in bacteria, making it possible to rapidly analyze samples even without prior purification of capsular polysaccharides.
AB - Polysialic acid capsules are the major virulence factors in Escherichia coli K1, K92, and groups B and C meningococci. The sialic acid monomers (2-keto-3-deoxy-5-acetamido-7,8,9- D - glycero - D - galacto -nonulosonic acids) comprising these homopolymeric polysaccharide chains can be selectively modified with 1,2- diamino-4,5-methylenedioxy-benzene to produce highly fluorescent quinoxalinone derivatives distinguished by their elution times during reverse phase chromatography. Here, we describe methods to release the constituent capsular polysialic acid monomers, and to detect and quantify them by sensitive fluorometry. There are relatively few 2-keto acids in bacteria, making it possible to rapidly analyze samples even without prior purification of capsular polysaccharides.
KW - Escherichia coli
KW - Fluorometry
KW - Polysialic acid capsules
KW - Release of sialic acid monomers
KW - Reverse phase chromatography
KW - Virulence factors
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U2 - 10.1007/978-1-62703-245-2_7
DO - 10.1007/978-1-62703-245-2_7
M3 - Chapter
C2 - 23299731
AN - SCOPUS:84879309256
SN - 9781627032445
T3 - Methods in Molecular Biology
SP - 109
EP - 120
BT - Bacterial Cell Surfaces
PB - Humana Press Inc.
ER -