22 February / March 2020
Figure 5: 50 pg/mL Extracted Insulin Aspart Standard Using bioZen MagBeads.
Figure 6: 500 pg/mL Extracted Insulin Aspart Standard Using bioZen MagBeads.
of the signature peptides is excellent which confirms a robust sample preparation protocol.
An important feature of this protocol is the similarity of the immunocapture procedure of rituximab compared to insulin. While the proteins are very different in structure, the immunocapture procedure had minimal modifications to achieve similar results. This fact suggests the broad applicability of the magnetic bead protocol and the ability to adopt it as a platform method.
Conclusion
As the complexity of therapeutic modalities increases, the bioanalytical field must adapt. Robust immunocapture and hybrid LBA/LC-
MS/MS protocols using bioZen MagBeads are demonstrated. Because hybrid LBA/LC-MSMS utilises MS as the detection, specificity of the assay can be improved while sensitivity can be similar to traditional ELISA methods.
By utilising streptavidin-coated magnetic beads, the method can be platformed in such a way that only the capture antibody or biotinylated target can be modified. This concept is demonstrated by the workflow being used for both a monoclonal antibody, as well as for a peptide therapeutic. Finally, this workflow not only extends to different therapeutic modalities, but it also applies to other large molecule bioanalytical workflows including intact quantitation or biotransformation, which also could implement a similar sample preparation strategy.
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Figure 7: XIC of the Signature Peptides of Rituximab on a bioZen Peptide XB-C18 column.
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