US 11,753,437 B2
Peptide purification using mixed-phase solid phase extraction material
Nils A. Kulak, Munich (DE); Garwin Pichler, Munich (DE); and Matthias Mann, Stockdorf (DE)
Assigned to MAX-PLANCK-GESELLSCHAFT ZUR FORDERUNG DER WISSENSCHAFTEN E.V., Munich (DE)
Appl. No. 15/741,804
Filed by MAX-PLANCK-GESELLSCHAFT ZUR FÖRDERUNG DER WISSENSCHAFTEN E.V., Munich (DE)
PCT Filed Jul. 8, 2016, PCT No. PCT/EP2016/066263
§ 371(c)(1), (2) Date Jan. 4, 2018,
PCT Pub. No. WO2017/005898, PCT Pub. Date Jan. 12, 2017.
Claims priority of application No. 15176142 (EP), filed on Jul. 9, 2015.
Prior Publication US 2018/0201644 A1, Jul. 19, 2018
Int. Cl. C07K 1/16 (2006.01); G01N 33/50 (2006.01)
CPC C07K 1/165 (2013.01) [G01N 33/5002 (2013.01)] 8 Claims
OG exemplary drawing
 
1. A method of purifying peptides and/or polypeptides bound to mixed- phase SPE material, said method comprising:
(a) loading a sample comprising peptides and/or polypeptides under acidic or neutral aqueous conditions on mixed-phase solid phase extraction (SPE) material comprising reversed phase material and/or ion exchange material;
(b) washing with
(ba) an acidic composition having a pH from 1 to 4 and comprising at least 50% (v/v) organic solvent; and
(bb) an acidic or neutral aqueous solution; and
(c) eluting the peptides and/or polypeptides from said mixed-phase SPE material with an alkaline composition comprising at least 50% (v/v) organic solvent;
wherein said mixed-phase SPE material is sulfonated poly-divinyl benzene (DVB) or sulfonated poly-styrene divinyl benzene (SDB); and
wherein step (a) is performed prior to step (b) which is performed prior to step (c).