Purification by centrifugal partition chromatography of amphiphilic compounds, glycolipids and pseudo-glycolipids synthesized by using cells.
Centrifugal partition chromatography (CPC) was applied to separate amphiphilic glycolipids and pseudo-glycolipids synthesized by using cells. Neutral and acidic lipid fractions were isolated by CPC under suitable conditions respectively. Separation of neutral lipid, Gb3-type and Gb4-type oligosaccharide synthesized by using cells, was performed with a two-phase solvent system composed of chloroform-methanol-water at a volume ratio of 5:6:4. On the other hand, separation of acidic lipid, GM3-type oligosaccharide synthesized by using cells, and ganglioside extracted from rat brain were performed with a two-phase solvent system composed of butanol-ethanol-1% acetic acid at a volume ratio of 4:1:5. 8.3mg of Gb3 analogue, 5.1mg of Gb4 analogue, and 19.5mg of GM3 analogue were purified from 3.2l of culture medium obtained by incubation of African green-monkey kidney (Vero) cells with 50 microM n-dodecyl beta-lactoside using CPC.
Kato
Tomohisa
T
Japan Chemical Innovation Institute, 1-3-5 Jimbocho, Chiyoda-ku, Tokyo 101-0051, Japan.
Miyagawa
Atsushi
A
Kasuya
Maria Carmelita Z
MC
Ito
Ayaka
A
Hatanaka
Kenichi
K
D000818:Animals; D001921:Brain; D045744:Cell Line, Tumor; D002522:Chlorocebus aethiops; D002845:Chromatography / Q000379:methods; D005679:G(M3) Ganglioside / Q000032:analysis / Q000302:isolation & purification*; D005732:Gangliosides / Q000032:analysis / Q000302:isolation & purification*; D006017:Glycolipids / Q000032:analysis / Q000302:isolation & purification*; D051379:Mice; D009844:Oligosaccharides / Q000032:analysis / Q000302:isolation & purification*; D051381:Rats; D014709:Vero Cells
D005679:G(M3) Ganglioside; D005732:Gangliosides; D006017:Glycolipids; D009844:Oligosaccharides