The Characterization of Drug-to-Antibody Ratio and Isoforms of ADCs Using LC and MS Technologies
| العنوان: | The Characterization of Drug-to-Antibody Ratio and Isoforms of ADCs Using LC and MS Technologies |
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| المؤلفون: | Weibin Chen, Thomas J. Porter, Robert E. Birdsall, Henry Shion, Frank W. Kotch, Reiko Iizuka, April Xu |
| المصدر: | YAKUGAKU ZASSHI. 137:525-528 |
| بيانات النشر: | Pharmaceutical Society of Japan, 2017. |
| سنة النشر: | 2017 |
| مصطلحات موضوعية: | 0301 basic medicine, Pharmacology, Gene isoform, Chromatography, Reverse-Phase, Antibody-drug conjugate, Immunoconjugates, Chromatography, Chemistry, Hydrophilic interaction chromatography, 010401 analytical chemistry, Antibodies, Monoclonal, Pharmaceutical Science, 01 natural sciences, Mass Spectrometry, 0104 chemical sciences, 03 medical and health sciences, 030104 developmental biology, Liquid chromatography–mass spectrometry, Mass spectrum, Protein Isoforms, Cysteine, Hydrophobic and Hydrophilic Interactions, Drug to antibody ratio, Chromatography, Liquid |
| الوصف: | Hydrophobic interaction chromatography (HIC) and MS are leading techniques for the characterization of the critical quality attributes (CQA) of antibody-drug conjugates (ADCs). This includes the average drug-to-antibody ratio (DAR) and drug loading distribution. A workflow that effectively utilizes the synergy between chromatography and detection technologies has been developed and was assessed using cysteine-conjugated ADCs. The DAR of low, moderate and high drug-loaded ADC samples were calculated from the chromatographic peak areas using LC(HIC)/UV or the deconvoluted mass spectra using native LC(SEC)/MS. The results of DAR by both technologies produced comparable results. In addition, the 2D-LC/MS system has been evaluated in combination with HIC and reversed-phase chromatography for structural identification. Individual peaks from the 1st dimension of the HIC separation were isolated online and re-directed to the 2nd dimension reversed-phase column. ADC was detected as the sub-units by MS and the conjugation site was identified via a middle down approach. |
| تدمد: | 1347-5231 0031-6903 |
| DOI: | 10.1248/yakushi.16-00255-1 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::2baf8da3fab64cc83a0dddaae82483d5 https://doi.org/10.1248/yakushi.16-00255-1 |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....2baf8da3fab64cc83a0dddaae82483d5 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13475231 00316903 |
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| DOI: | 10.1248/yakushi.16-00255-1 |