A radiochemical study of the irreversible adsorption of proteins on commercial reversed-phase HPLC packing materials is reported. The conditions of study are similar to those used in HPLC separation of protein. The effects of the amount and contact time of two proteins, ovalbumin and cytochrome c, are reported. Additional results include the effect of column pretreatment with protein, silanophilic mobile-phase blocking agent, and type of packing material on the extent of irreversible adsorption. The loss process is shown to be at least biphasic and the mechanisms of loss distinct for different proteins.
Bibliographical noteFunding Information:
This work was supported in part by a grant from the National Science Foundation. The authors also thank the 3M Company (St. Paul. Minn.) for their gracious cooperation in providing instrumentation and counting facilities for this work.
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