Epitope Identification and Affinity Determination of an Inhibiting Human Antibody to Interleukin IL8 (CXCL8) by SPR- Biosensor–Mass Spectrometry Combination

2019 ◽  
Vol 31 (1) ◽  
pp. 109-116 ◽  
Author(s):  
Pascal Wiegand ◽  
Loredana Lupu ◽  
Nico Hüttmann ◽  
Julia Wack ◽  
Stephan Rawer ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Human Antibody ◽  
Epitope Identification ◽  
Spr Biosensor

Antibody Epitope and Affinity Determination of the Myocardial Infarction Marker Myoglobin by SPR-Biosensor Mass Spectrometry

2020 ◽  
Vol 32 (1) ◽  
pp. 106-113
Author(s):  
Delia Mihoc ◽  
Loredana-Mirela Lupu ◽  
Pascal Wiegand ◽  
Wolfgang Kleinekofort ◽  
Oliver Müller ◽  
...  
Keyword(s):  
Myocardial Infarction ◽  
Mass Spectrometry ◽  
Spr Biosensor ◽  
Antibody Epitope

scholarly journals Identification and Affinity Determination of Protein-Antibody and Protein-Aptamer Epitopes by Biosensor-Mass Spectrometry Combination

2021 ◽  
Vol 22 (23) ◽  
pp. 12832
Author(s):  
Loredana-Mirela Lupu ◽  
Pascal Wiegand ◽  
Daria Holdschick ◽  
Delia Mihoc ◽  
Stefan Maeser ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Protein Interactions ◽  
Polyclonal Antibodies ◽  
Binding Constants ◽  
Dna Aptamer ◽  
Enzyme Replacement ◽  
Spr Biosensor ◽  
Wide Range ◽  
Antibody Complex

Analytical methods for molecular characterization of diagnostic or therapeutic targets have recently gained high interest. This review summarizes the combination of mass spectrometry and surface plasmon resonance (SPR) biosensor analysis for identification and affinity determination of protein interactions with antibodies and DNA-aptamers. The binding constant (KD) of a protein–antibody complex is first determined by immobilizing an antibody or DNA-aptamer on an SPR chip. A proteolytic peptide mixture is then applied to the chip, and following removal of unbound material by washing, the epitope(s) peptide(s) are eluted and identified by MALDI-MS. The SPR-MS combination was applied to a wide range of affinity pairs. Distinct epitope peptides were identified for the cardiac biomarker myoglobin (MG) both from monoclonal and polyclonal antibodies, and binding constants determined for equine and human MG provided molecular assessment of cross immunoreactivities. Mass spectrometric epitope identifications were obtained for linear, as well as for assembled (“conformational”) antibody epitopes, e.g., for the polypeptide chemokine Interleukin-8. Immobilization using protein G substantially improved surface fixation and antibody stabilities for epitope identification and affinity determination. Moreover, epitopes were successfully determined for polyclonal antibodies from biological material, such as from patient antisera upon enzyme replacement therapy of lysosomal diseases. The SPR-MS combination was also successfully applied to identify linear and assembled epitopes for DNA–aptamer interaction complexes of the tumor diagnostic protein C-Met. In summary, the SPR-MS combination has been established as a powerful molecular tool for identification of protein interaction epitopes.

Precise determination of 14 REEs in GSJ/AIST geochemical reference materials JCp-1 (coral) and JCt-1 (giant clam) using isotope dilution ICP-quadrupole mass spectrometry

2018 ◽  
Vol 52 (1) ◽  
pp. 75-79 ◽  
Author(s):  
Tsuyoshi Tanaka ◽  
Seung-Gu Lee ◽  
Taehoon Kim ◽  
Seunghee Han ◽  
Hyo Min Lee ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Reference Materials ◽  
Isotope Dilution ◽  
Precise Determination ◽  
Giant Clam ◽  
Quadrupole Mass ◽  
Quadrupole Mass Spectrometry ◽  
Geochemical Reference Materials
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