Fast protein liquid chromatography scale-up procedures for the preparation of low-molecular-weight proteins from urine

Abstract

A system for the rapid isolation of low molecular weight proteins from urine has been devised, illustrated by α 1-microglobulin, β 2-microglobulin, retinol binding protein, lysozyme and monoclonal light chains. Urine proteins from patients with tubular dysfunction were concentrated, either by ultrafiltration or ammonium sulphate precipitation. This was followed by gel chromatography on Sephadex G-50. The appropriate fractions were then separated by chromatography on Pharmacia monobead columns. A Mono Q strong anion exchanger was used for β 2-microglobulin, retinol binding protein, α 1-microglobulin free monoclonal light chains. Lysozyme was separated on a Mono S cation exchanger. The chromatography was first optimized on HR 5/5 columns and then scaled up to HR 16/10 columns.