PMID- 17947995
OWN - NLM
STAT- MEDLINE
DCOM- 20080122
LR  - 20080324
IS  - 1750-2799 (Electronic)
IS  - 1750-2799 (Linking)
VI  - 2
IP  - 10
DP  - 2007
TI  - Frontal affinity chromatography: sugar-protein interactions.
PG  - 2529-37
AB  - Frontal affinity chromatography using fluorescence detection (FAC-FD) is a 
      versatile technique for the precise determination of dissociation constants (Kd) 
      between glycan-binding proteins (lectins) and fluorescent-labeled glycans. A 
      series of glycan-containing solutions is applied to a lectin-immobilized column, 
      and the elution profile of each glycan (termed the 'elution front', V) is 
      compared with that (V0) for an appropriate control. Here we describe our standard 
      protocol using an automated FAC system (FAC-1), consisting of two isocratic 
      pumps, an autosampler, a column oven and two miniature columns connected to a 
      fluorescence detector. Analysis time for 100 sugar-protein interactions is 
      approximately 10 h, using as little as 2.5 pmol of pyridylaminated (PA) 
      oligosaccharide per analysis. Using FAC-FD, we have so far obtained quantitative 
      interaction data of >100 lectins for >100 PA oligosaccharides.
FAU - Tateno, Hiroaki
AU  - Tateno H
AD  - Research Center for Medical Glycoscience, National Institute of Advanced 
      Industrial Science and Technology, AIST Tsukuba Central 2, 1-1-1 Umezono, Ibaraki 
      305-8568, Japan.
FAU - Nakamura-Tsuruta, Sachiko
AU  - Nakamura-Tsuruta S
FAU - Hirabayashi, Jun
AU  - Hirabayashi J
LA  - eng
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - England
TA  - Nat Protoc
JT  - Nature protocols
JID - 101284307
RN  - 0 (Lectins)
RN  - 0 (Polysaccharides)
SB  - IM
MH  - Chromatography, Affinity/instrumentation/*methods
MH  - Lectins/chemistry/*metabolism
MH  - Polysaccharides/chemistry/*metabolism
EDAT- 2007/10/20 09:00
MHDA- 2008/01/23 09:00
CRDT- 2007/10/20 09:00
PHST- 2007/10/20 09:00 [pubmed]
PHST- 2008/01/23 09:00 [medline]
PHST- 2007/10/20 09:00 [entrez]
AID - nprot.2007.357 [pii]
AID - 10.1038/nprot.2007.357 [doi]
PST - ppublish
SO  - Nat Protoc. 2007;2(10):2529-37. doi: 10.1038/nprot.2007.357.