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 μm 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.