Chiral separation of T3 enantiomers using stereoselective antibodies as a selector in micro-HPLC.
This work deals with the application of stereoselective antibodies against L-T3 as a tailor-made chiral selector in micro-HPLC. The separations were performed in microbore columns using commercially available anti-L-T3 antibodies chemically bonded to 5 microm silica gel. The enantiomers of T3 were baseline separated under mild continuous isocratic elution conditions using 10 mM phosphate buffer, pH 7.4. The D-enantiomer eluted with the void volume, while the L-enantiomer was retained by the antibody phase and eluted second. An indirect competitive and non-competitive enzyme linked immunosorbent assay (ELISA) was used for testing the stereoselectivity of anti-L-T3 antibodies.
Koidl
Julia
J
Institute of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, Karl-Franzens-University, Universitätsplatz 1, A-8010 Graz, Austria.
Hödl
Heike
H
Schmid
Martin G
MG
Konrad
Marlene
M
Petschauer
Sabine
S
Kostner
Gerhard M
GM
Gübitz
Gerald
G
D000918:Antibody Specificity; D002846:Chromatography, Affinity; D002851:Chromatography, High Pressure Liquid; D004797:Enzyme-Linked Immunosorbent Assay; D008832:Microchemistry; D013237:Stereoisomerism; D014284:Triiodothyronine / Q000737:chemistry / Q000276:immunology / Q000302:isolation & purification*
D014284:Triiodothyronine