Size-exclusion chromatography of protected synthetic polypeptide tandems in an organic solvent.
A novel method for analytical and preparative size exclusion chromatography of large water-insoluble protected peptides in an organic solvent was developed. This method was applied to analysis and separation of protected synthetic peptide tandem repeats and to a control of the reaction of peptide fragment coupling. Columns containing Toyopearl HW-40, HW-50; HW-55 and HW-60 gels of Fine grade were used, and the selectivity of each sorbent, as well as the chromatographic behaviour of the peptides on them, were examined. Separation ranges of these gels applied to separation of protected peptides in DMF were shown to be much smaller (ca. 400-14,000 Da) than those of the same gels applied to protein separations in water buffers (100-1,000,000 Da). An initial evaluation of efficiency of Toyopearl HW Superfine grade gels with respect to similar separations was performed.
Karnoup
A S
AS
Institute of Protein Research, Russian Academy of Sciences, Pushchino, Moscow Region, Russia. karnup@sun.ipr.serpukhov.su
Shiryaev
V M
VM
Medvedkin
V N
VN
Mitin
Y V
YV
D000596:Amino Acids / Q000737:chemistry; D002138:Calibration; D002850:Chromatography, Gel / Q000379:methods*; D004121:Dimethyl Sulfoxide; D004126:Dimethylformamide; D005449:Fluorenes / Q000737:chemistry; D008970:Molecular Weight; D010455:Peptides / Q000032:analysis* / Q000302:isolation & purification; D011108:Polymers / Q000737:chemistry; D012995:Solubility; D012997:Solvents
D000596:Amino Acids; D005449:Fluorenes; C016456:N(alpha)-fluorenylmethyloxycarbonylamino acids; D010455:Peptides; D011108:Polymers; D012997:Solvents; C080256:Toyopearl HW 40; D004126:Dimethylformamide; D004121:Dimethyl Sulfoxide