Improved Recovery of a Radlolabeled Peptide with an Albumin-Treated Reversed-Phase HPLC Column

Reversed-phase high-performance liquid chromatography (RP-HPLC) is an important tool in the purification of radiolabeled peptides and proteins for immunoassay. However, for some proteins and peptides it is difficult to achieve reproducible behavior in RP-HPLC because of the low recovery of these compounds. Factors that can be varied to improve recovery include the strength or pH of the mobile phase, the chain length and spacing of groups on the reversed-phase support, and the flow rate or steepness of the elution gradient (1-5). ... In summary, we obtained better recovery and more reproducible chromatographic behavior for labeled 1-34 PTHrP with an albumin-pretreated reversed-phase column than with a new, untreated reversed-phase column. This treatment is simple and inexpensive and should be generally applicable for use in the purification of other radiolabeled peptides and proteins