24 May / June 2017 Conclusion
A universal LBA/LC-MS/MS workflow using high capacity streptavidin coated magnetic beads and reagents supplied with the BioBA sample prep kit enable the rapid development of pharmacokinetic assays of antibody based therapeutics. The versatility of the immunoenrichment strategy enables customisation by binding the appropriate biotin conjugated immunocapture reagent to the streptavidin coated beads. As shown here, the binding of the magnetic beads with a universal capture reagent followed by LC-MS/MS analysis enabled the accelerated
development of the total antibody assay for ado-trastuzumab emtansine with wide dynamic range, high selectivity, and high sensitivity, with an LLOQ at low nanogram levels. Microflow LC enabled up to 5x lower LLOQ vs. traditional flow LC providing a solution for applications where quantification is needed at low concentrations and/ or when sample volumes are limited. The workflow is generally applicable to mAb based therapeutics and results in faster assay development with more selectivity and lower LLOQs than typically achieved with LC-MS/ MS using a direct plasma or pellet digest.
References
[1] X. Geng, X. Kong, H. Hu, J. Chen, F. Yang, H. Liang, X. Chen, and Y. Hu, Hum Vaccin Immunother. 11(12) (2015) 2769–2776.
[2] D. Kuystermans and M. Al-Rubeai in M. Al-Rubeai (Editor), Animal Cell Culture, Springer, Dordrecht, The Netherlands, 2015, 717-57.
[3] M. Stern, R. Herrmann, Crit Rev Oncol Hematol. 54 (2005) 11-29.
[4]
https://en.wikipedia.org/wiki/List_of_ therapeutic_monoclonal_antibodies
New Range of Immobilised Chiral LC Columns
Nacalai Tesque have introduced COSMOSIL CHiRAL, a new range of immobilised polysaccharide LC chiral columns. Available in both 3µm and 5µm particle sizes, these immobilised chiral phases are compatible with both normal and reversed-phase conditions and are resistant to strong solvents that may be unsuitable for use with coated phases. Three different phases are currently available, CHiRAL A, CHiRAL B and CHiRAL C, in lengths of 150mm and 250mm and i.d.s of 4.6, 10 and 20mm. These columns are competitively priced and offer good chiral performance.
For more information or a technical catalogue contact authorised distributor Hichrom Ltd at
technical@hichrom.co.uk
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