BACKGROUND Detection limit challenges associated with measuring low-abundance protein biomarkers can be addressed with hybrid immunoaffinity-mass spectrometric assays, such as antipeptide antibody capture followed by liquid chromatography/tandem mass spectrometry (LC-MS/MS). Popular assay formats use magnetic bead-based immunoaffinity enrichment and nanoflow LC-MS/MS or high-flow immunoaffinity...

Affinity chromatography is a high resolution, high capacity, and one of the most powerful and diverse methods for separating proteins and other biological molecules of interest on the basis of a highly specific, reversible biological interaction between two molecules: an affinity ligand attached to a solid matrix to create a stationary phase, and a target molecule in a mobile phase. Specificall...

Highly efficient capillary electrochromatographic separations of cardiac glycosides and other steroids are presented. Employing butyl-derivatized silica particles as stationary phase resulted in a nearly three times faster electroosmotic flow (EOF) compared to capillary electrochromatography (CEC) with octadecyl silica particles. On-column focusing with a preconcentration factor of 180 was perf...

BACKGROUND Streptokinase is a potent activator of plasminogen to plasmin, the enzyme that can solubilize the fibrin network in blood clots. Streptokinase is currently used in clinical medicine as a thrombolytic agent. It is naturally secreted by β-hemolytic streptococci. METHODS To reach an efficient method of purification, an immunoaffinity chromatography method was developed that could puri...

We describe a simple, rapid immunoaffinity procedure for purifying the MB isoenzyme of creatine kinase. Immunoaffinity gel is prepared by linking a monoclonal antibody ("Conan-MB"), specific for this isoenzyme, to Sepharose 4B. Heart tissue is homogenized and fractionated with 40-70% saturated ammonium sulfate before it is applied to the immunoaffinity gel. CK-MB activity, retained on the gel, ...

Because of concern about possible transmission of ochratoxin A (OA) from contaminated grain adjuncts, development of a sensitive method for its determination in beer was investigated. Solid phase extraction (SPE) on C-18 and silica gel columns in series and on an immunoaffinity column (OchraTest) were used to obtain extracts for quantitation by reverse phase liquid chromatography with fluoresce...