Optimizing Hybrid LC–MS/MS Binding Conditions is Critical: Impact of Biotransformation on Quantification of Trastuzumab

Liu, Luna; Xu, Keyang; Li, Jenny; Maia, Mauricio; Mathieu, Mary; Elliott, Rebecca; Yang, Jihong; Nijem, Ihsan; Kaur, Surinder

doi:10.4155/bio-2018-0196

Cited by 18 publications

(18 citation statements)

References 24 publications

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“…Trastuzumab serum concentrations were determined by a validated high-performance liquid chromatography with tandem mass spectrometry (LC–MS/MS) detection described previously [ 15 ]. An affinity capture approach using streptavidin magnetic beads coupled with biotinylated recombinant human HER2 extracellular domain was used to enrich trastuzumab from human serum.…”

Section: Methodsmentioning

confidence: 99%

Pharmacokinetic and exploratory exposure–response analysis of pertuzumab in patients with operable HER2-positive early breast cancer in the APHINITY study

Kirschbrown¹,

Kågedal²,

Wang³

et al. 2019

Cancer Chemother Pharmacol

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Purpose To characterize the pharmacokinetics (PK) of, and perform an exploratory exposure–response (E–R) analysis for, pertuzumab in patients with HER2-positive early breast cancer (EBC) within the APHINITY study (NCT01358877, BIG 4–11/BO25126/TOC4939G). Methods A previously developed pertuzumab two-compartment linear population pharmacokinetic (popPK) model was subjected to external validation to examine appropriateness for describing pertuzumab concentrations from the APHINITY study. Pharmacokinetic drug–drug interactions (DDIs) between pertuzumab, trastuzumab, and chemotherapy were assessed by comparing observed serum or plasma C max , C min , and AUC last geometric mean ratios with 90% CIs. Predictions of pertuzumab C max,ss , C min,ss , and AUC ss were derived from individual parameter estimates and used in an exploratory E–R analysis. Results Using data from 72 patients, based on goodness-of-fit, the popPK model was deemed appropriate for predictions of individual exposures for subsequent comparisons to historical data, assessment of DDIs, and E–R analyses. No evidence of DDIs for pertuzumab on trastuzumab, trastuzumab on pertuzumab, or pertuzumab on chemotherapy PK was observed. Analyses of differences in exposure between patients with and without invasive disease-free survival events did not indicate improved efficacy with increased exposure. Overall Grade ≥ 3 diarrhea prevalence was higher with pertuzumab versus placebo, but was not greater with increasing pertuzumab exposure. No apparent E–R relationship was suggested with respect to other grade ≥ 3 AEs. Conclusion Overall, the limited available data from this exploratory study suggest that no dose adjustments are needed for pertuzumab when administered in combination with trastuzumab and an EBC chemotherapy regimen. Electronic supplementary material The online version of this article (10.1007/s00280-019-03826-1) contains supplementary material, which is available to authorized users.

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Section: Methodsmentioning

confidence: 99%

Pharmacokinetic and exploratory exposure–response analysis of pertuzumab in patients with operable HER2-positive early breast cancer in the APHINITY study

Kirschbrown¹,

Kågedal²,

Wang³

et al. 2019

Cancer Chemother Pharmacol

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“…Serum concentrations of trastuzumab were determined by a validated high-performance liquid chromatography assay with tandem mass spectrometry detection (minimum quantifiable concentration of 100 ng/mL) [13]. This assay showed acceptable accuracy (% difference) and % CV with ranges of − 8.08 to − 1.47% and 3.07 to 8.44%, respectively.…”

Section: Methodsmentioning

confidence: 99%

Pharmacokinetic and exposure–response analysis of pertuzumab in patients with HER2-positive metastatic gastric or gastroesophageal junction cancer

Kirschbrown¹,

Wang²,

Nijem³

et al. 2019

Cancer Chemother Pharmacol

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Purpose To characterize the pharmacokinetics (PK) of pertuzumab and trastuzumab in patients with HER2-positive metastatic gastric or gastroesophageal junction cancer in the randomized, double-blind, phase III JACOB study (NCT01774786), and to evaluate the appropriateness of the pertuzumab regimen in these patients. Methods Patients received 840 mg intravenous pertuzumab or placebo plus trastuzumab q3w and chemotherapy. Pertuzumab and trastuzumab were administered until disease progression or unacceptable toxicity. Chemotherapy was administered for up to six cycles or disease progression or unacceptable toxicity. Serum concentrations of pertuzumab and trastuzumab were measured. Pertuzumab PK was characterized across treatment cycles. The impact of anti-drug antibodies (ADAs) on pertuzumab PK and the impact of pertuzumab on trastuzumab PK were assessed. An exploratory exposure–efficacy analysis was also conducted. Results In total, 374 patients in the pertuzumab arm had evaluable PK data. The mean observed pertuzumab steady-state serum trough (minimum) concentration ( C min,ss ) ± standard deviation was 114 ± 51.8 μg/mL. The target pertuzumab C min,ss of ≥ 20 μg/mL was reached in 99.3% of patients at Cycle 5 (steady state) and beyond. Greater than 90% of patients were above the PK target right after the first pertuzumab dose. There was no apparent impact of ADAs on pertuzumab PK nor of pertuzumab on trastuzumab PK. There were no differences in overall survival across Cycle 1 pertuzumab ( C min ) or Cycle 5 pertuzumab ( C min,ss ) exposure quartiles. Conclusions Pertuzumab exposure in JACOB was consistent with prior studies in advanced gastric cancer and breast cancer. The 840 mg q3w dose allowed the majority of patients in JACOB to achieve target pertuzumab concentrations and appears to be an appropriate dose selection. Electronic supplementary material The online version of this article (10.1007/s00280-019-03871-w) contains supplementary material, which is available to authorized users.

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“…31 Deamidation of IYPTNGYTR can also occur in vivo and has been demonstrated to hamper the functionality of trastuzumab, most likely due to a conformational change that affects antigen binding. 18,31 Consistent evidence is lacking on whether surrogate peptides that reflect the total trastuzumab concentration (e.g., FTISADTSK, DTYIHWVR) or surrogate peptides more reflective of in vivo modifications (e.g., IYPTNGYTR, IYPTDGYTR) should be used. 15,16,18,31 Considering the small differences in mass (1 Da) and polarity between the IYPTNGYTR peptide and its deamidated form, the current method cannot resolve the two forms, and is therefore more reflective of the total trastuzumab concentration.…”

Section: Discussionmentioning

confidence: 99%

“…Essential for minimizing technical variations during sample preparation is the addition of an internal standard (IS), displaying similar or identical physical and chemical features, which allows monitoring of the full sample preparation workflow.- 10,12,14 LC-MS/MS methods using multiple reaction monitoring (MRM) for quantification of only trastuzumab in human serum have been described previously, including a method employing two surrogate peptides reaching a sensitivity of 5 μg/mL, and another reaching a sensitivity of 20 ng/mL with the latter using high resolution MS detection. [15][16][17][18] To facilitate quantification of co-administered mAbs, multiplex LC-MS/MS methods have been developed. However, until recently these assays always required specific reagents for affinity purification or specific stable isotope labeled (SIL) ISs.…”

Section: Introductionmentioning

confidence: 99%

“… 10 , 12 , 14 LC-MS/MS methods using multiple reaction monitoring (MRM) for quantification of only trastuzumab in human serum have been described previously, including a method employing two surrogate peptides reaching a sensitivity of 5 μg/mL, and another reaching a sensitivity of 20 ng/mL with the latter using high resolution MS detection. 15 – 18 …”

Section: Introductionmentioning

confidence: 99%

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Development and validation of an LC-MS/MS method for simultaneous quantification of co-administered trastuzumab and pertuzumab

Schokker

Bonardi

Molenaar

et al. 2020

mAbs

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Given the increasing use of combination therapy with multiple monoclonal antibodies (mAbs), there is a clinical need for multiplexing assays. For the frequently co-administered anti-human epidermal growth factor receptor 2 (HER2) mAbs trastuzumab and pertuzumab, we developed a high-throughput and robust hybrid ligand-binding liquid chromatography-mass spectrometry (LC-MS)/MS quantitative assay. Nanomolar concentrations of trastuzumab and pertuzumab were determined in 10 µL serum samples after extraction by affinity purification through protein A beads, followed by on-bead reduction, alkylation, and trypsin digestion. After electrospray ionization, quantification was obtained by multiple reaction monitoring LC-MS/MS using SILuMab as an internal standard. The method was validated according to the current guidelines from the US Food and Drug Administration and the European Medicines Agency. Assay linearity was established in the ranges 0.250-250 μg/mL for trastuzumab and 0.500-500 μg/mL for pertuzumab. The method was accurate and selective for the simultaneous determination of trastuzumab and pertuzumab in clinical samples, thereby overcoming the limitation of ligand binding assays that cannot quantify mAbs targeting the same receptor. Furthermore, this method requires a small blood volume, which reduces blood collection time and stress for patients. The assay robustness was verified in a clinical trial where trastuzumab and pertuzumab concentrations were determined in 670 serum samples. As we used commercially available reagents and standards, the described generic bioanalytical strategy can easily be adapted to multiplex quantifications of other mAb combinations in non-clinical and clinical samples.

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Optimizing Hybrid LC–MS/MS Binding Conditions is Critical: Impact of Biotransformation on Quantification of Trastuzumab

Cited by 18 publications

References 24 publications

Pharmacokinetic and exploratory exposure–response analysis of pertuzumab in patients with operable HER2-positive early breast cancer in the APHINITY study

Pharmacokinetic and exploratory exposure–response analysis of pertuzumab in patients with operable HER2-positive early breast cancer in the APHINITY study

Pharmacokinetic and exposure–response analysis of pertuzumab in patients with HER2-positive metastatic gastric or gastroesophageal junction cancer

Development and validation of an LC-MS/MS method for simultaneous quantification of co-administered trastuzumab and pertuzumab

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