Purification of radiolabeled and native polypeptides by gel permeation high-performance liquid chromatography

✍ Scribed by C. Lazure; M. Dennis; J. Rochemont; N.G. Seidah; M. Chrétien

• Publisher: Elsevier Science
• Year: 1982
• Tongue: English
• Weight: 743 KB
• Volume: 125
• Category: Article
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(82)90022-7

✦ Synopsis

Some results and observations concerning the use of protein columns are presented. The combined use of four protein columns having different fractionation ranges together with a volatile triethylamine formate buffer allowed the sieving of various polypeptides according to their molecular weights over a range of 500 to 150,000. The addition of 4 or 6 M guanidine-HCl permitted the reduction of aggregation with no sacrifice in resolution or linearity. With that denaturant, rapid separation, and molecular weight determination in the range 500-90,000 is easily accomplished. Moreover, sample recoveries as determined with radiolabeled proteins always exceeded 70% while radioimmunoassay techniques can be directly applied to the column eluate. Applications to quick identification of natural fragments of a serine protease, tonin, analysis of maturation products of pro-opiomelanocortin in an in vitro pulse experiment and finally quantitation by radioimmunoassays of pituitary peptides and elution of their 'r51labeled derivatives are described.

' This paper was presented at the International Symposium on HPLC of Proteins and Peptides,