Rapid separation of biologically active Syrian hamster lymphotoxin in high yield by size-exclusion high-performance liquid chromatography.

A rapid, simple and reproducible high-performance liquid chromatography (HPLC) procedure is described for the partial purification of biologically active Syrian hamster lymphotoxin. Lymphotoxin, a series of lymphocyte derived immunological glycoprotein hormones with molecular weights of 20,000 to 56,000, has been partially purified by a single-step HPLC procedure or by a two-step isoelectric focusing-HPLC procedure. The HPLC separation method uses silica-based protein separation columns eluted with a 0.1 M sodium phosphate, pH 7.4, 0.1% polyethylene glycol 4000 buffer at a flow-rate of 0.5 ml/min at room temperature. Nearly complete recoveries of biologically active lymphotoxin can be obtained with as much as a 13-fold purification in less than 1 h.