A method of purifying a peptide
A technology for cyclic peptides and purity, applied in the field of purified peptides
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Embodiment 1
[0113] A 66% pure eptifibatide TFA salt prepared by solid-phase synthesis was used for purification on a polymer-based resin-packed column. Eptifibatide TFA salt was initially dissolved in a 1:1 mixture of acetonitrile and 50 mM acetic acid to obtain a clear solution. The obtained solution was further diluted to an acetonitrile concentration of 5% and an eptifibatide concentration of <2 g / L using 50 mM acetic acid. The solution was filtered and loaded onto the column.
[0114] Amberchrom HPR10 (particle size 10 μm and pore size ) resin-packed column. The filtered eptifibatide solution was loaded onto the column at a flow rate < 360 cm / h. Loading of the peptide on the column was done at <10 g / L concentration of resin. After loading, the column was rinsed with a lower percentage (5%) of acetonitrile in 50 mM acetic acid solution. Pure product was eluted from the column by performing a linear gradient of 8-14% of acetonitrile (buffer B) for 25 CV, while 50 mM acetic acid wa...
Embodiment 2
[0120] A 69.5% pure eptifibatide TFA salt prepared by solid phase synthesis was used for purification on a polymer-based resin packed column. Eptifibatide TFA salt was initially dissolved in a 1:1 mixture of acetonitrile and 50 mM acetic acid to obtain a clear solution. The obtained solution was further diluted to an acetonitrile concentration of 5% and an eptifibatide concentration of <2 g / L using 50 mM acetic acid. The solution was filtered and loaded onto the column. The pH of sodium acetate was adjusted to 3.0 with acetic acid.
[0121] Amberchrom HPR10 (particle size 10 μm and pore size ) resin-packed column. The filtered solution of eptifibatide was loaded onto the column at a flow rate < 360 cm / h. Loading of the peptide on the column was performed at a concentration < 10 g / L of the resin. After loading, the column was rinsed with a lower percentage (5%) of acetonitrile in 10 mM sodium acetate, pH 3.0. Pure product was eluted from the column by performing a 9-12% ...
Embodiment 3
[0125] A 69.5% pure eptifibatide TFA salt prepared by solid phase synthesis was used for purification on a polymer-based resin packed column. Eptifibatide TFA salt was initially dissolved in a 1:1 mixture of acetonitrile and 50 mM acetic acid to obtain a clear solution. The obtained solution was further diluted to an acetonitrile concentration of 5% and an eptifibatide concentration of <2 g / L using 50 mM acetic acid. The solution was filtered and loaded onto the column.
[0126] Amberchrom HPR10 (particle size 10 μm and pore size ) resin-packed column. The filtered eptifibatide solution was loaded onto the column at a flow rate < 360 cm / h. Loading of the peptide on the column was performed at a concentration < 10 g / L of the resin. After loading, the column was rinsed with a lower percentage (5%) of acetonitrile in 10 mM citric acid, pH 2.5. Pure product was eluted from the column by performing a 9-12% linear gradient of acetonitrile (buffer B) for 25CV, while 10 mM citri...
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