Set-up of large laboratory-scale chromatographic separations of poly(ethylene glycol) derivatives of the growth hormone-releasing factor 1-29 analogue.
In this paper we report the scale-up of the purification of poly(ethylene glycol) (PEG) derivatives of the growth hormone-releasing factor 1-29, from laboratory scale (100 mg of bulk starting material) to larger scale (3 g of bulk), through the use of a cation-exchange TSK-SP-5PW chromatographic column. A one-step purification process capable of purifying large amounts of mono-PEGylated GRF species from the crude reaction mixture was developed. A simple, straightforward stepwise gradient elution separation was developed at laboratory scale and then scaled up with a larger column packed with a chromatographic resin with the same chemistry which maintained the laboratory-scale separation profile. Active material recovery and material purity remained constant through the scale-up from the 13-microm stationary phase to the 25-microm larger column. Overall, the gram GRF equivalent/batch process scale showed to be quite reproducible, and could be considered as a good platform for scale up to production scale.
Piquet
G
G
Istituto di Ricerca Cesare Serono, Drug Delivery Systems, Bioconjugates development and scale-up department, BioIndustry Park del Canavese, Turin, Italy. gilles.piquet@serono.com
Gatti
M
M
Barbero
L
L
Traversa
S
S
Caccia
P
P
Esposito
P
P
D002411:Cation Exchange Resins; D002852:Chromatography, Ion Exchange / Q000379:methods*; D013007:Growth Hormone-Releasing Hormone / Q000737:chemistry / Q000302:isolation & purification*; D010446:Peptide Fragments / Q000737:chemistry / Q000302:isolation & purification*; D011092:Polyethylene Glycols / Q000737:chemistry*; D015203:Reproducibility of Results
D002411:Cation Exchange Resins; D010446:Peptide Fragments; D011092:Polyethylene Glycols; D013007:Growth Hormone-Releasing Hormone; C059148:somatotropin releasing hormone (1-29)