Sponsor
This work was supported in parts by grants from the National Institutes of Health (CA-115531 and RR-02584) and the Robert A. Welch Foundation (AT-584).
Published In
Bioorganic & Medicinal Chemistry
Document Type
Post-Print
Publication Date
2011
Subjects
Chelates -- Synthesis, Contrast media (Diagnostic imaging), Magnetic resonance imaging, Monoclonal antibodies, Serum albumin
Abstract
Reactive surface lysine groups on the chimeric monoclonal antibody (3G4) and on human serum albumin (HSA) were labeled with two different PARACEST chelates. Between 7.4 – 10.1 chelates were added per 3G4 molecule and between 5.6 – 5.9 chelates per molecule of HSA, depending upon which conjugation chemistry was used. The immunoreactivity of 3G4 as measured by ELISA assays was highly dependent upon the number of attached chelates: 88% immunoreactivity with 7.4 chelates per antibody versus only 17% immunoreactivity with 10.1 chelates per antibody. Upon conjugation to 3G4, the bound water lifetime of Eu-1 increased only marginally, up from 53 μs for the non-conjugated chelate to 65–77 μs for conjugated chelates. Conjugation of a chelate Eu-2 to HSA via a single side-chain group also resulted in little or no change in bound water lifetime (73–75 μs for both the conjugated and non-conjugated forms). These data indicate that exchange of water molecules protons between the inner-sphere site on covalently attached PARACEST agent and bulk water is largely unaffected by the mode of attachment of the agent to the protein and likely its chemical surroundings on the surface of the protein.
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DOI
10.1016/j.bmc.2010.06.026
Persistent Identifier
https://archives.pdx.edu/ds/psu/32391
Citation Details
Published as: Vasalatiy, O., Zhao, P., Woods, M., Marconescu, A., Castillo-Muzquiz, A., Thorpe, P., Kiefer, G. E., & Sherry, A. D. (2011). Strategies for labeling proteins with PARACEST agents. Bioorganic & medicinal chemistry, 19(3), 1106–1114. https://doi.org/10.1016/j.bmc.2010.06.026
Description
This is the author’s version of a work that was accepted for publication in Bioorganic & Medicinal Chemistry. Changes resulting from the publishing process, such as peer review, editing, corrections, structural formatting, and other quality control mechanisms may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. A definitive version was subsequently published in Bioorganic & Medicinal Chemistry, 2011 Feb 1;19(3):1106-14. doi: 10.1016/j.bmc.2010.06.026.
© 2011. This manuscript version is made available under the CC-BY-NC-ND 4.0 license http://creativecommons.org/licenses/by-nc-nd/4.0/
*At the time of publication Mark Woods was affiliated with the University of Texas at Dallas