Mapping the interactions of selective biochemical probes of antibody conformation by hydrogen-deuterium exchange mass spectrometry
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Mapping the interactions of selective biochemical probes of antibody conformation by hydrogen-deuterium exchange mass spectrometry. / Leurs, Ulrike; Beck, Hermann; Bonnington, Lea; Lindner, Ingo; Pol, Ewa; Rand, Kasper Dyrberg.
In: ChemBioChem, Vol. 18, No. 11, 01.06.2017, p. 1016-1021.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Mapping the interactions of selective biochemical probes of antibody conformation by hydrogen-deuterium exchange mass spectrometry
AU - Leurs, Ulrike
AU - Beck, Hermann
AU - Bonnington, Lea
AU - Lindner, Ingo
AU - Pol, Ewa
AU - Rand, Kasper Dyrberg
N1 - © 2017 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.
PY - 2017/6/1
Y1 - 2017/6/1
N2 - Protein-based pharmaceuticals represent the fastest growing group of drugs in development in the pharmaceutical industry. One of the major challenges in the discovery, development and distribution of biopharmaceuticals is the assessment of changes in their higher-order structure due to chemical modifications. Here, we investigate the interactions of three different biochemical probes (Fabs) generated to detect conformational changes in a therapeutic IgG1 antibody (mAbX) by local hydrogen-deuterium exchange mass spectrometry (HDX-MS). We show that two of the probes target the Fc part of the antibody, while the third probe binds to the hinge region. Through HDX-ETD, we can distinguish specific binding patterns of the Fc-binding probes on mAbX at the single amino acid level. Preliminary surface plasmon resonance (SPR) experiments show that these domain-selective Fab probes are sensitive to conformational changes in distinct regions of a full-length therapeutic antibody upon oxidation.
AB - Protein-based pharmaceuticals represent the fastest growing group of drugs in development in the pharmaceutical industry. One of the major challenges in the discovery, development and distribution of biopharmaceuticals is the assessment of changes in their higher-order structure due to chemical modifications. Here, we investigate the interactions of three different biochemical probes (Fabs) generated to detect conformational changes in a therapeutic IgG1 antibody (mAbX) by local hydrogen-deuterium exchange mass spectrometry (HDX-MS). We show that two of the probes target the Fc part of the antibody, while the third probe binds to the hinge region. Through HDX-ETD, we can distinguish specific binding patterns of the Fc-binding probes on mAbX at the single amino acid level. Preliminary surface plasmon resonance (SPR) experiments show that these domain-selective Fab probes are sensitive to conformational changes in distinct regions of a full-length therapeutic antibody upon oxidation.
KW - Journal Article
U2 - 10.1002/cbic.201600670
DO - 10.1002/cbic.201600670
M3 - Journal article
C2 - 28346764
VL - 18
SP - 1016
EP - 1021
JO - ChemBioChem
JF - ChemBioChem
SN - 1439-4227
IS - 11
ER -
ID: 177084155