Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic Islets
An immunoencapsulation strategy for pancreatic islets aimed to reduce the risk of rejection in transplanted patients due to the immune response of the host organism is proposed. In this sense, a polyethylene glycol (PEG) hydrogel functionalized with an immunosuppressive antibody (Ab), such as Cytoto...
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MDPI AG
2020-09-01
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| Series: | Applied Sciences |
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| Online Access: | https://www.mdpi.com/2076-3417/10/17/6056 |
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| author | Anna Cavallo Ugo Masullo Alessandra Quarta Alessandro Sannino Amilcare Barca Tiziano Verri Marta Madaghiele Laura Blasi |
| author_facet | Anna Cavallo Ugo Masullo Alessandra Quarta Alessandro Sannino Amilcare Barca Tiziano Verri Marta Madaghiele Laura Blasi |
| author_sort | Anna Cavallo |
| collection | DOAJ |
| description | An immunoencapsulation strategy for pancreatic islets aimed to reduce the risk of rejection in transplanted patients due to the immune response of the host organism is proposed. In this sense, a polyethylene glycol (PEG) hydrogel functionalized with an immunosuppressive antibody (Ab), such as Cytotoxic T-lymphocyte antigen-4 Ig (CTLA4-Ig), would act as both passive and active barrier to the host immune response. To demonstrate the feasibility of this approach, a photopolymerizable-PEG was conjugated to the selected antibody and the PEG-Ab complex was used to coat the islets. Moreover, to preserve the antigen-recognition site of the antibody during the conjugation process, a controlled immobilization method was setup through the attachment of the His-tagged antigen to a solid support. In detail, a gold-coated silicon wafer functionalized with 11-Mercaptoundecanoic acid was used as a substrate for further modification, leading to a nickel(II)-terminated ligand surface. Then, the immobilized antigen was recognized by the corresponding antibody that was conjugated to the PEG. The antibody-PEG complex was detached from the support prior to be photopolymerized around the islets. First, this immobilization method has been demonstrated for the green fluorescent protein (GFP)–anti-green fluorescent protein (Anti-GFP) antigen-antibody pair, as proof of principle. Then, the approach was extended to the immunorelevant B7-1 CTLA-4-Ig antigen-antibody pair, followed by the binding of Acryl-PEG to the immobilized constant region of the antibody. In both cases, after using an elution protocol, only a partial recovery of the antibody-PEG complex was obtained. Nevertheless, the viability and the functional activity of the encapsulated islets, as determined by the glucose-stimulated insulin secretion (GSIS) assay, showed the good compatibility of this approach. |
| first_indexed | 2024-03-10T16:39:35Z |
| format | Article |
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| institution | Directory Open Access Journal |
| issn | 2076-3417 |
| language | English |
| last_indexed | 2024-03-10T16:39:35Z |
| publishDate | 2020-09-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Applied Sciences |
| spelling | doaj.art-39bf05b1d0c240508f657f088d72e6b22023-11-20T12:08:42ZengMDPI AGApplied Sciences2076-34172020-09-011017605610.3390/app10176056Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic IsletsAnna Cavallo0Ugo Masullo1Alessandra Quarta2Alessandro Sannino3Amilcare Barca4Tiziano Verri5Marta Madaghiele6Laura Blasi7Department of Engineering for Innovation, University of Salento, Via Monteroni, 73100 Lecce, ItalyDepartment of Engineering for Innovation, University of Salento, Via Monteroni, 73100 Lecce, ItalyCNR Nanotec, Institute of Nanotechnology, via Monteroni, 73100 Lecce, ItalyDepartment of Engineering for Innovation, University of Salento, Via Monteroni, 73100 Lecce, ItalyDepartment of Biological and Environmental Sciences and Tecnologies, University of Salento, 73100 Lecce, ItalyDepartment of Biological and Environmental Sciences and Tecnologies, University of Salento, 73100 Lecce, ItalyDepartment of Engineering for Innovation, University of Salento, Via Monteroni, 73100 Lecce, ItalyCNR Nanotec, Institute of Nanotechnology, via Monteroni, 73100 Lecce, ItalyAn immunoencapsulation strategy for pancreatic islets aimed to reduce the risk of rejection in transplanted patients due to the immune response of the host organism is proposed. In this sense, a polyethylene glycol (PEG) hydrogel functionalized with an immunosuppressive antibody (Ab), such as Cytotoxic T-lymphocyte antigen-4 Ig (CTLA4-Ig), would act as both passive and active barrier to the host immune response. To demonstrate the feasibility of this approach, a photopolymerizable-PEG was conjugated to the selected antibody and the PEG-Ab complex was used to coat the islets. Moreover, to preserve the antigen-recognition site of the antibody during the conjugation process, a controlled immobilization method was setup through the attachment of the His-tagged antigen to a solid support. In detail, a gold-coated silicon wafer functionalized with 11-Mercaptoundecanoic acid was used as a substrate for further modification, leading to a nickel(II)-terminated ligand surface. Then, the immobilized antigen was recognized by the corresponding antibody that was conjugated to the PEG. The antibody-PEG complex was detached from the support prior to be photopolymerized around the islets. First, this immobilization method has been demonstrated for the green fluorescent protein (GFP)–anti-green fluorescent protein (Anti-GFP) antigen-antibody pair, as proof of principle. Then, the approach was extended to the immunorelevant B7-1 CTLA-4-Ig antigen-antibody pair, followed by the binding of Acryl-PEG to the immobilized constant region of the antibody. In both cases, after using an elution protocol, only a partial recovery of the antibody-PEG complex was obtained. Nevertheless, the viability and the functional activity of the encapsulated islets, as determined by the glucose-stimulated insulin secretion (GSIS) assay, showed the good compatibility of this approach.https://www.mdpi.com/2076-3417/10/17/6056polyethylene glycolimmunoisolationpancreatic islets |
| spellingShingle | Anna Cavallo Ugo Masullo Alessandra Quarta Alessandro Sannino Amilcare Barca Tiziano Verri Marta Madaghiele Laura Blasi Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic Islets Applied Sciences polyethylene glycol immunoisolation pancreatic islets |
| title | Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic Islets |
| title_full | Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic Islets |
| title_fullStr | Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic Islets |
| title_full_unstemmed | Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic Islets |
| title_short | Design of Antibody-Functionalized Polymeric Membranes for the Immunoisolation of Pancreatic Islets |
| title_sort | design of antibody functionalized polymeric membranes for the immunoisolation of pancreatic islets |
| topic | polyethylene glycol immunoisolation pancreatic islets |
| url | https://www.mdpi.com/2076-3417/10/17/6056 |
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