Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method
Trastuzumab emtansine (T-DM1, brand name: Kadcyla<sup>®</sup>) is the first FDA-approved antibody-drug conjugate (ADC) for metastatic human epidermal growth factor receptor 2 positive (HER2+) breast cancer. It consists of three components: trastuzumab, an anti-HER2 monoclonal antibody, m...
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MDPI AG
2021-10-01
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| Online Access: | https://www.mdpi.com/2076-3417/11/20/9437 |
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| author | Seo-jin Park Byeong ill Lee Min-Ho Park Jangmi Choi Yuri Park Min-jae Park Jeong-hyeon Lim Jiyu Lee Sangsoo Hwang Jeongmin Lee Young G. Shin |
| author_facet | Seo-jin Park Byeong ill Lee Min-Ho Park Jangmi Choi Yuri Park Min-jae Park Jeong-hyeon Lim Jiyu Lee Sangsoo Hwang Jeongmin Lee Young G. Shin |
| author_sort | Seo-jin Park |
| collection | DOAJ |
| description | Trastuzumab emtansine (T-DM1, brand name: Kadcyla<sup>®</sup>) is the first FDA-approved antibody-drug conjugate (ADC) for metastatic human epidermal growth factor receptor 2 positive (HER2+) breast cancer. It consists of three components: trastuzumab, an anti-HER2 monoclonal antibody, maytansinoid (DM1) as a cytotoxic drug, and maleimidomethyl cyclohexane-1-carboxylate (MCC) as a linker. In particular, the MCC linker is known as a non-cleavable linker and has a feature of being conjugated to DM1 by a covalent thioether bond. In this study, we developed an immuno-affinity capture liquid chromatography-mass spectrometric (LC-MS/MS) assay for quantifying the antibody-conjugated drug (acDrug) component of T-DM1. To quantify acDrug, desulfurated DM1 was prepared using a chemical desulfuration pretreatment and quantified as an acDrug. A quadratic regression (weighted 1/concentration), with equation y = ax<sup>2</sup> + bx + c, was used to fit the calibration curves over the concentration range of 17.09~1709.44 ng/mL for the acDrug of T-DM1. The quantification run met the in-house acceptance criteria of ±25% accuracy and precision values for the quality control (QC) samples. In conclusion, an immuno-affinity capture LC-MS/MS assay was successfully developed to quantify acDrug of T-DM1 and applied to evaluate in vitro plasma linker stability and preclinical pharmacokinetic (PK) study in rats. This assay could be helpful when applied to other ADCs with the same linker-cytotoxic drug platform. |
| first_indexed | 2024-03-10T06:44:56Z |
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| institution | Directory Open Access Journal |
| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-03-10T06:44:56Z |
| publishDate | 2021-10-01 |
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| series | Applied Sciences |
| spelling | doaj.art-066a6efbfb3247288f5d3129c8490d402023-11-22T17:18:35ZengMDPI AGApplied Sciences2076-34172021-10-011120943710.3390/app11209437Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric MethodSeo-jin Park0Byeong ill Lee1Min-Ho Park2Jangmi Choi3Yuri Park4Min-jae Park5Jeong-hyeon Lim6Jiyu Lee7Sangsoo Hwang8Jeongmin Lee9Young G. Shin10Institute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaInstitute of Drug Research and Development, College of Pharmacy, Chungnam National University, Daejeon 34134, KoreaTrastuzumab emtansine (T-DM1, brand name: Kadcyla<sup>®</sup>) is the first FDA-approved antibody-drug conjugate (ADC) for metastatic human epidermal growth factor receptor 2 positive (HER2+) breast cancer. It consists of three components: trastuzumab, an anti-HER2 monoclonal antibody, maytansinoid (DM1) as a cytotoxic drug, and maleimidomethyl cyclohexane-1-carboxylate (MCC) as a linker. In particular, the MCC linker is known as a non-cleavable linker and has a feature of being conjugated to DM1 by a covalent thioether bond. In this study, we developed an immuno-affinity capture liquid chromatography-mass spectrometric (LC-MS/MS) assay for quantifying the antibody-conjugated drug (acDrug) component of T-DM1. To quantify acDrug, desulfurated DM1 was prepared using a chemical desulfuration pretreatment and quantified as an acDrug. A quadratic regression (weighted 1/concentration), with equation y = ax<sup>2</sup> + bx + c, was used to fit the calibration curves over the concentration range of 17.09~1709.44 ng/mL for the acDrug of T-DM1. The quantification run met the in-house acceptance criteria of ±25% accuracy and precision values for the quality control (QC) samples. In conclusion, an immuno-affinity capture LC-MS/MS assay was successfully developed to quantify acDrug of T-DM1 and applied to evaluate in vitro plasma linker stability and preclinical pharmacokinetic (PK) study in rats. This assay could be helpful when applied to other ADCs with the same linker-cytotoxic drug platform.https://www.mdpi.com/2076-3417/11/20/9437antibody-drug conjugate (ADC)antibody-conjugated drug (acDrug)quantificationLC-MS/MSbioanalysisnon-cleavable linker |
| spellingShingle | Seo-jin Park Byeong ill Lee Min-Ho Park Jangmi Choi Yuri Park Min-jae Park Jeong-hyeon Lim Jiyu Lee Sangsoo Hwang Jeongmin Lee Young G. Shin Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method Applied Sciences antibody-drug conjugate (ADC) antibody-conjugated drug (acDrug) quantification LC-MS/MS bioanalysis non-cleavable linker |
| title | Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method |
| title_full | Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method |
| title_fullStr | Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method |
| title_full_unstemmed | Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method |
| title_short | Quantification for Antibody-Conjugated Drug in Trastuzumab Emtansine and Application to In Vitro Linker Stability and In Vivo Pharmacokinetic Study in Rat Using an Immuno-Affinity Capture Liquid Chromatography-Mass Spectrometric Method |
| title_sort | quantification for antibody conjugated drug in trastuzumab emtansine and application to in vitro linker stability and in vivo pharmacokinetic study in rat using an immuno affinity capture liquid chromatography mass spectrometric method |
| topic | antibody-drug conjugate (ADC) antibody-conjugated drug (acDrug) quantification LC-MS/MS bioanalysis non-cleavable linker |
| url | https://www.mdpi.com/2076-3417/11/20/9437 |
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