A smart membrane based on an antigen-responsive hydrogel.
Hydrogel membranes have been fabricated that incorporate antibody/antigen moieties. The permeability of large solutes through these membranes is dependent on the presence of soluble antigen that can compete with the internal interactions between antibody and antigen leading to an increase in gel mesh size. Specifically, the membrane's structure is based on a dextran backbone grafted with a fluorescein isothiocyanate (FITC) antigen and a sheep anti-FITC IgG antibody. The backbone is covalently cross-linked by conjugated divinyl sulfone (DVS) groups. The gel structure is additionally stabilized by affinity crosslinks formed by biospecific interactions between the bound IgG and FITC. FTIR spectra of the gel are consistent with formation of covalent bonds between cysteine groups in the IgG and DVS groups in the dextran. Results obtained using isothermal titration calorimetry (ITC) confirmed the competitive interaction binding between IgG-FITC-dextran and free sodium fluorescein at pH 5.0. Scanning electron microscopy (SEM) of samples prepared using cryofixation and cryofracturing techniques showed that observed changes in permeability correlate with free fluorescein-dependent structural changes in the gel. Three-dimensional images obtained from confocal laser scanning microscopy show that these changes occur throughout the gel and indicate that SEM results are not artifacts of sample preparation. The permeability of these gels, as shown by blue-dextran (12 kDa) diffusion, increases in response to the presence of free fluorescein of the external medium, which causes competitive displacement of the affinity cross-links. Sequential addition and removal of sodium fluorescein showed that these permeability changes are reversible.
Zhang
Rongsheng
R
Department of Chemical Engineering, University of Bath, Claverton Down, Bath BA2 7AY, UK.
Bowyer
Adrian
A
Eisenthal
Robert
R
Hubble
John
J
D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D000941:Antigens / Q000276:immunology* / Q000378:metabolism*; D001672:Biocompatible Materials / Q000737:chemistry; D002151:Calorimetry / Q000379:methods; D003432:Cross-Linking Reagents / Q000737:chemistry; D003545:Cysteine / Q000737:chemistry; D003911:Dextrans / Q000737:chemistry / Q000648:ultrastructure; D004058:Diffusion; D019793:Fluorescein; D016650:Fluorescein-5-isothiocyanate; D005456:Fluorescent Dyes; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D006863:Hydrogen-Ion Concentration; D021621:Imaging, Three-Dimensional; D007074:Immunoglobulin G / Q000737:chemistry / Q000378:metabolism* / Q000648:ultrastructure; D008567:Membranes, Artificial; D018613:Microscopy, Confocal; D015394:Molecular Structure; D010539:Permeability; D011485:Protein Binding; D017550:Spectroscopy, Fourier Transform Infrared; D013450:Sulfones / Q000737:chemistry
D000906:Antibodies; D000941:Antigens; D001672:Biocompatible Materials; D003432:Cross-Linking Reagents; D003911:Dextrans; D005456:Fluorescent Dyes; D007074:Immunoglobulin G; D008567:Membranes, Artificial; D013450:Sulfones; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate; C009873:divinyl sulfone; D016650:Fluorescein-5-isothiocyanate; D003545:Cysteine; D019793:Fluorescein