LC-MS/MS Validation Analysis of Trastuzumab Using dSIL Approach for Evaluating Pharmacokinetics
Rohit H. Budhraja,
Milin A. Shah,
Mahendra Suthar,
Arun Yadav,
Sahil P. Shah,
Prashant Kale,
Parisa Asvadi,
Mariadhas Valan Arasu,
Naif Abdullah Al-Dhabi,
Chun Geon Park,
Young-Ock Kim,
Hak Jae Kim,
Y. K. Agrawal,
Ravi. K. Krovidi
Affiliations
Rohit H. Budhraja
Panomics Lambda Therapeutic Research Limited, Gota, Ahmedabad, Gujarat 382481, India
Milin A. Shah
Panomics Lambda Therapeutic Research Limited, Gota, Ahmedabad, Gujarat 382481, India
Mahendra Suthar
Panomics Lambda Therapeutic Research Limited, Gota, Ahmedabad, Gujarat 382481, India
Arun Yadav
Panomics Lambda Therapeutic Research Limited, Gota, Ahmedabad, Gujarat 382481, India
Sahil P. Shah
Panomics Lambda Therapeutic Research Limited, Gota, Ahmedabad, Gujarat 382481, India
Prashant Kale
Panomics Lambda Therapeutic Research Limited, Gota, Ahmedabad, Gujarat 382481, India
Parisa Asvadi
Analytical Development and Global Regulatory Affairs, Intas Pharmaceuticals Ltd., Ahmedabad, Gujarat 382213, India
Mariadhas Valan Arasu
Department of Botany and Microbiology, Addiriyah Chair for Environmental Studies, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia
Naif Abdullah Al-Dhabi
Department of Botany and Microbiology, Addiriyah Chair for Environmental Studies, College of Science, King Saud University, P.O. Box 2455, Riyadh 11451, Saudi Arabia
Chun Geon Park
Department of Medicinal Crop Research, Rural Development Administration, Eumseong, Chungbuk 369-873, Korea
Young-Ock Kim
Department of Medicinal Crop Research, Rural Development Administration, Eumseong, Chungbuk 369-873, Korea
Hak Jae Kim
Department of Clinical Pharmacology, College of Medicine, Soonchunhyang University, Cheonan 31151, Korea
Y. K. Agrawal
Department of Pharmaceutical Sciences, Gujarat Forensic Sciences University, Gandhinagar 382481, India
Ravi. K. Krovidi
Panomics Lambda Therapeutic Research Limited, Gota, Ahmedabad, Gujarat 382481, India
Quantitative targeted proteomics based approaches deploy state-of-the-art Liquid chromatography tandem mass spectrometry LC-MS technologies and are evolving as a complementary technique to standard ligand-binding based assays. Advancements in MS technology, which have augmented the specificity, selectivity and sensitivity limits of detection and freedom from antibody generation, have made it amicable towards various clinical applications. In our current work, a surrogate peptide based quantitative proteomics assessment is performed by selecting specific signature peptides from the complementary determining region CDR region of trastuzumab (Herclon®, Roche products in India). We developed a double Stable Isotope Label (dSIL) approach by using two different surrogate peptides to evaluate the proteolytic digestion efficiency and accurate quantification of the target analyte peptide of Herclon® in human serum. Method validation experiments were meticulously performed as per bioanalytical method validation guidelines. The dSIL approach, using an LC-MS/MS based quantification assay demonstrated good linearity over a range of 5–500 µg/mL of Herclon®, and validation experimental data is in compliance with bioanalytical regulatory guidelines.