Evaluation and separation of steroid-bovine serum albumin conjugates by high-performance liquid chromatography.
The conventional methods for characterization of steroid immunogens are based on the determination of the total amount of hapten bound to the protein carrier either by the UV spectroscopy or titration of unsubstituted amino groups. These methods do not allow more detailed insight into the immunogen composition. HPLC of the immunogen combined with UV detection is a relatively rapid and convenient method enabling determination of the hapten content in each fraction and, eventually, separation of individual fractions differing in the hapten content or purification of crude product.
Hill
M
M
Institute of Endocrinology, Prague, Czech Republic.
Lapcík
O
O
Hampl
R
R
D000097:Acetonitriles; D002851:Chromatography, High Pressure Liquid; D003687:Dehydroepiandrosterone / Q000031:analogs & derivatives / Q000302:isolation & purification; D004958:Estradiol / Q000031:analogs & derivatives / Q000302:isolation & purification; D006241:Haptens / Q000737:chemistry / Q000302:isolation & purification; D010091:Oximes / Q000302:isolation & purification; D012710:Serum Albumin, Bovine / Q000302:isolation & purification*; D012997:Solvents; D013256:Steroids / Q000302:isolation & purification*; D013739:Testosterone / Q000031:analogs & derivatives / Q000302:isolation & purification; D014612:Vaccines / Q000737:chemistry / Q000302:isolation & purification
D000097:Acetonitriles; D006241:Haptens; D010091:Oximes; D012997:Solvents; D013256:Steroids; D014612:Vaccines; C106648:dehydroepiandrosterone-7-O-(carboxymethyl)oxime; C067733:testosterone-3-carboxymethyloxime; D012710:Serum Albumin, Bovine; C002912:estradiol-6-(O-carboxymethyl)oxime; D013739:Testosterone; D003687:Dehydroepiandrosterone; D004958:Estradiol; C032159:acetonitrile