期刊
PROTEOMICS
卷 7, 期 10, 页码 1624-1633出版社
WILEY
DOI: 10.1002/pmic.200600778
关键词
host cell proteins; impurity analysis; ligand library; mass spectrometry; peptide ligands
Capture and amplification of low-level contaminants in purified preparations of recombinant DNA products is described here in the case of mAb meant for human consumption. Such a process is based on treatment with a vastly heterogenous ligand library composed of hexapeptides bound to a polyhydroxymethacrylate resin. Upon this treatment, a protein solution is recovered with normalized relative concentration ratios, in which high-abundance proteins are strongly reduced and rare proteins are highly concentrated. Upon 2-D map analysis, the relatively few spots present in control monoclonals were seen to increase in number, reaching > 100 visible polypeptide chains in the pI/M-r plane. Most of these newly emerged spots were subjected to MS analysis and were found to be composed mainly of three classes of proteins: those derived from proteins present in the culture broth (notably albumin and transferrin), fragments of the desired final product, covering M-r ranges from as low as 5 up to 45 kDa and some aggregates of light and heavy chains of Igs (mostly dimers and trimers). This ligand library thus appears to be a formidable tool for exploring and bringing to the limelight the hidden proteome.
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