A radiochemical study of irreversible protein loss on high-performance liquid chromatography column frits

• Published in: Analytical biochemistry Vol. 144; no. 1; pp. 128 - 131
• Main Authors: Sadek, Paul C., Carr, Peter W., Bowers, Larry D., Haddad, Louis C.
• Format: Journal Article
• Language: English
• Published: San Diego, CA Elsevier Inc 1985; Elsevier
• Subjects: Adsorption; Analytical, structural and metabolic biochemistry; Biological and medical sciences; chromatography; Chromatography, High Pressure Liquid - instrumentation; frits; Fundamental and applied biological sciences. Psychology; General aspects, investigation methods; high-performance liquid chromatography; HPLC; protein adsorption; protein recovery; Proteins; Proteins - isolation & purification; radioactive labelling; radioiodination; Stainless Steel; Surface Properties; protein adsorption; stainless steel; proteins; chromatography; radioiodination; protein recovery; HPLC; frits; Proteins; High performance; Liquid chromatography; Purification; Separation; Adsorption
• ISSN: 0003-2697; 1096-0309
• DOI: 10.1016/0003-2697(85)90093-4

Much success has been achieved in the separation and purification of a wide range of proteins using various high-pressure liquid chromatography techniques. Quantitative analyses of proteins which require 100% mass recovery of the protein are still beset with problems, especially when the total injected amount of protein decreases to below 10 μg. Stainless-steel frits have been cited for their deleterious effects on chromatography in general. In addition, the frits have specifically been found to be a significant contributor to irreversible protein loss—particularly when protein sample sizes are on the order of 1 μg or less. The findings presented below should therefore be of concern to those using HPLC for protein work.