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Publication Detail
Homogeneous antibody fragment conjugation by disulfide bridging introduces 'spinostics'.
  • Publication Type:
    Journal article
  • Publication Sub Type:
    Journal Article
  • Authors:
    Schumacher FF, Sanchania VA, Tolner B, Wright ZV, Ryan CP, Smith ME, Ward JM, Caddick S, Kay CW, Aeppli G, Chester KA, Baker JR
  • Publication date:
    2013
  • Pagination:
    1525, ?
  • Journal:
    Sci Rep
  • Volume:
    3
  • Country:
    England
  • PII:
    srep01525
  • Language:
    eng
  • Keywords:
    Antibodies, Antigens, Disulfides, Electron Spin Resonance Spectroscopy, Humans, Maleimides, Spin Labels
Abstract
A major obstacle to the efficient production of antibody conjugates for therapy and diagnosis is the non-ideal performance of commonly used chemical methods for the attachment of effector-molecules to the antibody of interest. Here we demonstrate that this limitation can be simply addressed using 3,4-substituted maleimides to bridge and thus functionalize disulfide bonds to generate homogeneous antibody conjugates. This one-step conjugation reaction is fast, site-specific, quantitative and generates products with full binding activity, good plasma stability and the desired functional properties. Furthermore, the rigid nature of this modification by disulfide bridging enables the successful detection of antigen with a spin labeled antibody fragment by continuous-wave electron paramagnetic resonance (cw-EPR), which we report here for the first time. Antigen detection is concentration dependent, observable in human blood and allows the discrimination of fragments with different binding affinity. We envisage broad potential for antibody based in-solution diagnostic methods by EPR or 'spinostics'.
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