2018
DOI: 10.1039/c8an01178h
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Qualitative analysis of antibody–drug conjugates (ADCs): an experimental comparison of analytical techniques of cysteine-linked ADCs

Abstract: Antibody-drug conjugates (ADCs) are an emerging type of biotherapeutics that utilize multiple tissuespecific antibodies combined with a range of linker designs to enable the transportation and selective release of cytotoxic drugs in close proximity to tumours. Consisting of antibodies conjugated to small drug molecules through a variety of linkers, ADCs are chemically complex analytes. Here we present a unique experimental comparison of four techniques for ADC analysis: hydrophobic interaction chromatography (… Show more

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Cited by 30 publications
(34 citation statements)
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“…As has been reported previously [ 27 ], the average DAR value for a given ADC production batch may vary up to ± 0.3 DAR units between analytical techniques and a variability of 0.01–0.4 DAR units between different injections of the same ADC for a specific technique. For the triple quadrupole mass analyzer, DAR varied between injections up to 0.5 DAR units for one batch of ADCs, but was below 0.2 DAR units for all other batches, and on average within 0.32 DAR units from HIC-UV/Vis-derived values and was therefore in good agreement with previous findings.…”
Section: Resultssupporting
confidence: 55%
“…As has been reported previously [ 27 ], the average DAR value for a given ADC production batch may vary up to ± 0.3 DAR units between analytical techniques and a variability of 0.01–0.4 DAR units between different injections of the same ADC for a specific technique. For the triple quadrupole mass analyzer, DAR varied between injections up to 0.5 DAR units for one batch of ADCs, but was below 0.2 DAR units for all other batches, and on average within 0.32 DAR units from HIC-UV/Vis-derived values and was therefore in good agreement with previous findings.…”
Section: Resultssupporting
confidence: 55%
“…RP-HPLC analysis was also carried out for DAR determination ( Supplementary Materials Figures S9-S12), however slightly lower DAR results were obtained. It has been reported in a previous paper that the DAR value from RP-HPLC may be slightly lower than that measured from HIC analysis [10,20,23]. RP-HPLC analysis requires relatively high temperature and denaturing pretreatment with a combination of urea and DTT.…”
Section: Site-specific Conjugation and Purification Of Trastuzumab-ajmentioning
confidence: 77%
“…However, comparative studies regarding multiple types of analysis for DAR determination using a single ADC batch are extremely limited in a peer-reviewed setting. [8][9][10][11][12] Moreover, no examination to compare HPLC results among different ADCs prepared by various TCEP equivalents was reported to best of our knowledge. TCEP titration is the most crucial investigation to determine the appropriate equivalents to obtain cysteine-based ADCs with target DAR.…”
Section: Introductionmentioning
confidence: 99%