Publication

LC-MS/MS-based monitoring of in vivo protein biotransformation: quantitative determination of trastuzumab and its deamidation products in human plasma

Bults, P., Bischoff, R., Bakker, H., Gietema, J. A. & van de Merbel, N. C., 11-Feb-2016, In : Analytical Chemistry. 88, 3, p. 1871-1877 7 p.

Research output: Contribution to journalArticleAcademicpeer-review

APA

Bults, P., Bischoff, R., Bakker, H., Gietema, J. A., & van de Merbel, N. C. (2016). LC-MS/MS-based monitoring of in vivo protein biotransformation: quantitative determination of trastuzumab and its deamidation products in human plasma. Analytical Chemistry, 88(3), 1871-1877. https://doi.org/10.1021/acs.analchem.5b04276

Author

Bults, Peter ; Bischoff, Rainer ; Bakker, Hilde ; Gietema, Jourik A. ; van de Merbel, Nico C. / LC-MS/MS-based monitoring of in vivo protein biotransformation : quantitative determination of trastuzumab and its deamidation products in human plasma. In: Analytical Chemistry. 2016 ; Vol. 88, No. 3. pp. 1871-1877.

Harvard

Bults, P, Bischoff, R, Bakker, H, Gietema, JA & van de Merbel, NC 2016, 'LC-MS/MS-based monitoring of in vivo protein biotransformation: quantitative determination of trastuzumab and its deamidation products in human plasma', Analytical Chemistry, vol. 88, no. 3, pp. 1871-1877. https://doi.org/10.1021/acs.analchem.5b04276

Standard

LC-MS/MS-based monitoring of in vivo protein biotransformation : quantitative determination of trastuzumab and its deamidation products in human plasma. / Bults, Peter; Bischoff, Rainer; Bakker, Hilde; Gietema, Jourik A.; van de Merbel, Nico C.

In: Analytical Chemistry, Vol. 88, No. 3, 11.02.2016, p. 1871-1877.

Research output: Contribution to journalArticleAcademicpeer-review

Vancouver

Bults P, Bischoff R, Bakker H, Gietema JA, van de Merbel NC. LC-MS/MS-based monitoring of in vivo protein biotransformation: quantitative determination of trastuzumab and its deamidation products in human plasma. Analytical Chemistry. 2016 Feb 11;88(3):1871-1877. https://doi.org/10.1021/acs.analchem.5b04276


BibTeX

@article{facd43adf8ab4b96b28ecefa448208de,
title = "LC-MS/MS-based monitoring of in vivo protein biotransformation: quantitative determination of trastuzumab and its deamidation products in human plasma",
abstract = "An LC-MS/MS-based method is described for quantitatively monitoring the in vivo deamidation of the biopharmaceutical monoclonal antibody trastuzumab at a crucial position in its complementarity determining region (CDR). The multiplexed LC-MS/MS assay using selected reaction monitoring (SRM) allows simultaneous quantitation of five molecular species derived from trastuzumab after tryptic digestion: a stable signature peptide (FTISADTSK), a deamidation-sensitive signature peptide (IYPTNGYTR), its deamidated products (IYPTDGYTR and IYPTisoDGYTR) and a succinimide intermediate (IYPTsuccGYTR). Digestion of a 50-μL plasma sample is performed at pH 7 for three hours at 37°C, which combines a reasonable (>80%) digestion efficiency with a minimal (<1%) formation of deamidation products during digestion. Rapid in vitro deamidation was observed at higher pH, leading to a (large) overestimation of the concentrations of deamidation products in the original plasma sample. The LC-MS/MS method was validated in accordance with international bioanalytical guidelines over the clinically relevant range of 0.5 to 500 µg/mL with bias and CV values well below 15%. Deamidation of trastuzumab was observed in plasma both in a 56-day in vitro forced degradation study (up to 37% of the total drug concentration) and in samples obtained from breast cancer patients after treatment with the drug for several months (up to 25%). Comparison with a validated ELISA method for trastuzumab showed that deamidation of the drug at the CDR leads to a loss of recognition by the antibodies used in the ELISA assay.",
keywords = "MONOCLONAL-ANTIBODY, MASS-SPECTROMETRY, QUANTIFICATION, SERUM, MECHANISM, THERAPY",
author = "Peter Bults and Rainer Bischoff and Hilde Bakker and Gietema, {Jourik A.} and {van de Merbel}, {Nico C.}",
year = "2016",
month = feb,
day = "11",
doi = "10.1021/acs.analchem.5b04276",
language = "English",
volume = "88",
pages = "1871--1877",
journal = "Analytical Chemistry",
issn = "0003-2700",
publisher = "AMER CHEMICAL SOC INC",
number = "3",

}

RIS

TY - JOUR

T1 - LC-MS/MS-based monitoring of in vivo protein biotransformation

T2 - quantitative determination of trastuzumab and its deamidation products in human plasma

AU - Bults, Peter

AU - Bischoff, Rainer

AU - Bakker, Hilde

AU - Gietema, Jourik A.

AU - van de Merbel, Nico C.

PY - 2016/2/11

Y1 - 2016/2/11

N2 - An LC-MS/MS-based method is described for quantitatively monitoring the in vivo deamidation of the biopharmaceutical monoclonal antibody trastuzumab at a crucial position in its complementarity determining region (CDR). The multiplexed LC-MS/MS assay using selected reaction monitoring (SRM) allows simultaneous quantitation of five molecular species derived from trastuzumab after tryptic digestion: a stable signature peptide (FTISADTSK), a deamidation-sensitive signature peptide (IYPTNGYTR), its deamidated products (IYPTDGYTR and IYPTisoDGYTR) and a succinimide intermediate (IYPTsuccGYTR). Digestion of a 50-μL plasma sample is performed at pH 7 for three hours at 37°C, which combines a reasonable (>80%) digestion efficiency with a minimal (<1%) formation of deamidation products during digestion. Rapid in vitro deamidation was observed at higher pH, leading to a (large) overestimation of the concentrations of deamidation products in the original plasma sample. The LC-MS/MS method was validated in accordance with international bioanalytical guidelines over the clinically relevant range of 0.5 to 500 µg/mL with bias and CV values well below 15%. Deamidation of trastuzumab was observed in plasma both in a 56-day in vitro forced degradation study (up to 37% of the total drug concentration) and in samples obtained from breast cancer patients after treatment with the drug for several months (up to 25%). Comparison with a validated ELISA method for trastuzumab showed that deamidation of the drug at the CDR leads to a loss of recognition by the antibodies used in the ELISA assay.

AB - An LC-MS/MS-based method is described for quantitatively monitoring the in vivo deamidation of the biopharmaceutical monoclonal antibody trastuzumab at a crucial position in its complementarity determining region (CDR). The multiplexed LC-MS/MS assay using selected reaction monitoring (SRM) allows simultaneous quantitation of five molecular species derived from trastuzumab after tryptic digestion: a stable signature peptide (FTISADTSK), a deamidation-sensitive signature peptide (IYPTNGYTR), its deamidated products (IYPTDGYTR and IYPTisoDGYTR) and a succinimide intermediate (IYPTsuccGYTR). Digestion of a 50-μL plasma sample is performed at pH 7 for three hours at 37°C, which combines a reasonable (>80%) digestion efficiency with a minimal (<1%) formation of deamidation products during digestion. Rapid in vitro deamidation was observed at higher pH, leading to a (large) overestimation of the concentrations of deamidation products in the original plasma sample. The LC-MS/MS method was validated in accordance with international bioanalytical guidelines over the clinically relevant range of 0.5 to 500 µg/mL with bias and CV values well below 15%. Deamidation of trastuzumab was observed in plasma both in a 56-day in vitro forced degradation study (up to 37% of the total drug concentration) and in samples obtained from breast cancer patients after treatment with the drug for several months (up to 25%). Comparison with a validated ELISA method for trastuzumab showed that deamidation of the drug at the CDR leads to a loss of recognition by the antibodies used in the ELISA assay.

KW - MONOCLONAL-ANTIBODY

KW - MASS-SPECTROMETRY

KW - QUANTIFICATION

KW - SERUM

KW - MECHANISM

KW - THERAPY

U2 - 10.1021/acs.analchem.5b04276

DO - 10.1021/acs.analchem.5b04276

M3 - Article

C2 - 26713683

VL - 88

SP - 1871

EP - 1877

JO - Analytical Chemistry

JF - Analytical Chemistry

SN - 0003-2700

IS - 3

ER -

ID: 27138159