Development of a spectroscopic assay for bifunctional ligand-protein conjugates based on copper

Nuclear Medicine and Biology
Erik D BradyM W Brechbiel

Abstract

A simple, non-radioactive method for the determination of ligand-to-protein ratio (L/P) for novel ligand-antibody conjugates has been developed based on an exchange equilibrium with the purple Cu(II) complex of arsenazo III. The method requires a UV/Vis spectrometer and has been verified for monoclonal antibody Herceptin conjugates of a variety of ligand modalities, including common macrocyclic compounds NOTA and TETA, and with a new bifunctional tachpyridine (1H-Pyrrole-1-butanamide,N-[4-[[(1alpha,3alpha,5alpha)-3,5-bis[(2-pyridinylmethyl)amino]cyclohexyl](2-pyridinylmethyl)amino]butyl]-2,5-dihydro-2,5-dioxo-(9CI)). The spectroscopically derived values for L/P were verified by titration of the ligand-antibody conjugate with 64Cu. In each case, the value obtained by UV/Vis spectroscopy matches that found by radiolabeling. The method is rapid, taking less than 30 minutes with each ligand in this study.

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Citations

Jun 21, 2008·Bioconjugate Chemistry·Hyun-Soon ChongMartin W Brechbiel
Feb 4, 2014·Applied Radiation and Isotopes : Including Data, Instrumentation and Methods for Use in Agriculture, Industry and Medicine·Usha PandeyGrace Samuel
Jul 21, 2014·Annals of Nuclear Medicine·Mythili KameswaranGrace Samuel
Jun 30, 2012·Nuclear Medicine and Biology·Ravindra A De SilvaBuck E Rogers
Aug 17, 2011·Nuclear Medicine and Biology·Mojtaba SaloutiMohammad javad Rasaee
Nov 3, 2015·Applied Radiation and Isotopes : Including Data, Instrumentation and Methods for Use in Agriculture, Industry and Medicine·Mythili KameswaranGrace Samuel
Mar 22, 2016·The Indian Journal of Medical Research·Mythili KameswaranMeera Venkatesh
Dec 9, 2014·Contrast Media & Molecular Imaging·Charles TruilletOlivier Tillement
Jun 9, 2012·Dalton Transactions : an International Journal of Inorganic Chemistry·G MontavonA Morgenstern
Feb 10, 2017·Journal of Labelled Compounds & Radiopharmaceuticals·Mohini GuleriaSharmila Banerjee
Sep 19, 2007·Clinical Cancer Research : an Official Journal of the American Association for Cancer Research·Fares Al-EjehMichael P Brown
Apr 6, 2021·Journal of Labelled Compounds & Radiopharmaceuticals·Zbynek NovyFrantisek Trejtnar
Jun 12, 2010·Applied Radiation and Isotopes : Including Data, Instrumentation and Methods for Use in Agriculture, Industry and Medicine·Samira RasanehFariba Johari Daha

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