HPLC Application of Protein Variant

Side products from synthesis: The chromatogram for BMP-14 has several minor peaks eluting prior to the main peak. These are more apparent in the blown-up view below. Peaks in this location are generally deamidation variants. Peaks to the right of the main peak are generally oxidation variants or have disulfide bridges connecting the wrong residues. This chromatogram is typical of recombinant proteins in the TGF-beta family and growth factors in general

Cation-Exchange HPLC of BMP-14 (Bone Morphogenetic Protein 14)
Column: PolyCAT A™, 100×4.6mm; 3µ, 1000Å
Mobile Phase: A) 20mM K-PO4, pH 6.0, with 20% ACN; B) Same + 0.6M NaCl
Gradient: 20′ linear, 0-100%B, then 5′ at 100%B 1.0ml/min A220
Sample: 25µg BMP-14 in 20µl of (Mobile Phase A:10mM HCl = 2:1)

Column Name : PolyCAT A™ Columns

Compound Name : Bone Morphogenetic Protein 14