High performance liquid affinity chromatography (HPLAC) and its application to the separation of enzymes and antigens

High performance liquid chromatography (HPLC) has seen a spectacular development during the last few years due to its rapidity and its high resolving power [l-7] .Recently, HPLC has also been applied to include biological macromolecules such as polynucleotides, polysaccharides and proteins [3-71.This paper describes the first use of bioaffinity supports (in contrast to normally used 'non-biological' stationary phases) to the resolution of biological macromolecules in combination with the technique of HPLC, and for this novel technique we suggest the term: high performance liquid affinity chromatography, HPLAC.Examples are given of enzyme and isozyme separations using an immobilised general ligand, AMP, and of albumins employing an immobilised immunosorbent, anti-serum albumin. Materials and methodsCrystalline LADH (1.85 U/mg), #-NADH (grade III), ,&NAD (grade III), BSA (crystalline) and AMP were purchased from Sigma (St. Louis, MO.).LDH from pig heart (400 U/mg) and rabbit muscle (550 U/mg) were supplied by Boehringer (Mannheim,