کد مقاله | کد نشریه | سال انتشار | مقاله انگلیسی | نسخه تمام متن |
---|---|---|---|---|
1184356 | 1491798 | 2016 | 5 صفحه PDF | دانلود رایگان |
• The use of nESI-MS to examine intact Antibody Drug Conjugates (ADC).
• Use of TEAA as charge reducing agent improves cysteine-linked ADC characterization.
• TEAA preserves the intact mAb and facilitate easy drug load determination by native MS.
• This method is particularly beneficial for users of low resolution mass spectrometers.
Antibody-drug-conjugates (ADC) are a growing class of anticancer biopharmaceuticals. Conjugation of cysteine linked ADCs, requires initial reduction of mAb inter-chain disulfide bonds, as the drugs are attached via thiol chemistry. This results in the active mAb moiety being transformed from a covalently linked tetramer to non-covalently linked complexes, which hinders precise determination of drug load with LC–MS. Here, we show how the addition of the charge reducing agent triethylammonium acetate (TEAA) preserves the intact mAb structure, is well suited to the study of cysteine linked conjugates and facilitates easy drug load determination by direct infusion native MS.
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Journal: EuPA Open Proteomics - Volume 11, June 2016, Pages 23–27