Scale-up methodology for the preparative purification of peptide M.
An octadecapeptide, peptide M, the epitope of a retinal protein that induces experimental autoimmune uveitis, was synthesized and purified by preparative reversed-phase chromatography. The flow-rate and gradient conditions for maximum separation of impurities were determined on a 30 x 0.39 cm I.D. column of Delta Pak (15-microns spherical C18-bonded silica with 300-A pores). The maximum amount of peptide that was resolved under these conditions was then determined experimentally. Using a scale factor dependent on the square of the column diameters, the flow-rate and amount loaded were increased 164 times on a 30 x 5 cm I.D. column of the same packing. The same resolution was achieved. Batches of 200-342 mg were chromatographed with reproducible results, providing a total yield of 394 mg of pure peptide.
Knight
M
M
Peptide Technologies Corporation, Washington, DC 20017.
Strickler
M P
MP
Stone
M J
MJ
Chiodetti
L
L
Gluch
S
S
Shinohara
T
T
D002851:Chromatography, High Pressure Liquid / Q000379:methods; D005136:Eye Proteins / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000302:isolation & purification*
D005136:Eye Proteins; D010446:Peptide Fragments; C052615:peptide M, retinal S antigen