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Influence of Label and Charge Density on the Association of the Therapeutic Monoclonal Antibodies Trastuzumab and Cetuximab Conjugated to Anionic Fluorophores.
Pauli, Jutta; Pochstein, Marieke; Haase, Andrea; Napp, Joanna; Luch, Andreas; Resch-Genger, Ute.
Afiliação
  • Pauli J; Federal Institute for Materials Research and Testing (BAM), Division 1.10 Biophotonics, Richard-Willstaetter-Strasse 11, 12489, Berlin, Germany.
  • Pochstein M; Federal Institute for Materials Research and Testing (BAM), Division 1.10 Biophotonics, Richard-Willstaetter-Strasse 11, 12489, Berlin, Germany.
  • Haase A; German Federal Institute for Risk Assessment (BfR), Department of Chemical and Product Safety, Max-Dohrn-Strasse 8-10, 10589, Berlin, Germany.
  • Napp J; Institute of Interventional and Diagnostic Radiology, University Medical Center Göttingen, Robert-Koch-Strasse 40, 37075, Göttingen, Germany.
  • Luch A; Department of Haematology and Medical Oncology, University Medical Center Göttingen, Robert-Koch-Strasse 40, <postCode/37075, Göttingen, Germany.
  • Resch-Genger U; Department of Molecular Biology of Neuronal Signal, Max-Planck-Institute of Experimental Medicine, Hermann-Rein-Strasse 3, 37075, Göttingen, Germany.
Chembiochem ; 18(1): 101-110, 2017 Jan 03.
Article em En | MEDLINE | ID: mdl-27790811
The design of bright and functional dye-protein conjugates requires hydrophilic and stable fluorophores with high molar absorption coefficients and high fluorescence quantum yields, which must not be prone to dimerization, as well as conservation of protein function and suppression of protein association. Although many synthetic dyes meet these needs, the influence of dye charge on bioconjugate performance is commonly neglected. This encouraged us to assess the spectroscopic properties, antibody functionality, binding behavior, folding, and association of conjugates of the therapeutic antibodies trastuzumab and cetuximab with the red cyanine dyes S0586, S2381, and 6SIDCC (bearing two, three, and six sulfonate groups, respectively). Our results demonstrate a negligible effect of dye labeling on antibody folding, yet a strong influence of label charge and density on antibody isoelectric points and association. Especially 6SIDCC decreased strongly the isoelectric points of both antibodies and their heavy or light chains even at low labeling degrees, thus favoring protein association. Although an increasingly negative dye charge reduces antigen affinity as shown in a competitive immunoassay, all conjugates still bound to cells overexpressing the target of the respective antibody. Obviously, dyes that cause minimum dimerization with a small number of charged groups are best for conjugate brightness, minimum protein association, and strong target binding. This underlines the need to consider dye charge for the rational design of conjugates with optimum performance.
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Texto completo: 1 Base de dados: MEDLINE Assunto principal: Corantes Fluorescentes / Cetuximab / Trastuzumab Tipo de estudo: Risk_factors_studies Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article

Texto completo: 1 Base de dados: MEDLINE Assunto principal: Corantes Fluorescentes / Cetuximab / Trastuzumab Tipo de estudo: Risk_factors_studies Limite: Humans Idioma: En Ano de publicação: 2017 Tipo de documento: Article