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.
Bibliographical noteFunding Information:
We wish to thank the 3M Company for their support and the use of their facilities, the Separations Group for donating the Vydac packing material, and the National Science Foundation (CHE8217363) for support of this work.
- protein adsorption
- protein recovery
- stainless steel