Synthesis of protein conjugates adsorbed on cationic liposomes surface
The well-known Toll like receptor 9 (TLR9) agonist CpG ODN has shown promising results as vaccine adjuvant in preclinical and clinical studies, however its in vivo stability and potential systemic toxicity remain a concern. In an effort to overcome these issues, different strategies have been explor...
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2020-01-01
|
| Series: | MethodsX |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S221501612030162X |
| _version_ | 1818426625143865344 |
|---|---|
| author | Despo Chatzikleanthous Robert Cunliffe Filippo Carboni Maria Rosaria Romano Derek T. O'Hagan Craig W. Roberts Yvonne Perrie Roberto Adamo |
| author_facet | Despo Chatzikleanthous Robert Cunliffe Filippo Carboni Maria Rosaria Romano Derek T. O'Hagan Craig W. Roberts Yvonne Perrie Roberto Adamo |
| author_sort | Despo Chatzikleanthous |
| collection | DOAJ |
| description | The well-known Toll like receptor 9 (TLR9) agonist CpG ODN has shown promising results as vaccine adjuvant in preclinical and clinical studies, however its in vivo stability and potential systemic toxicity remain a concern. In an effort to overcome these issues, different strategies have been explored including conjugation of CpG ODN with proteins or encapsulation/adsorption of CpG ODN into/onto liposomes. Although these methods have resulted in enhanced immunopotency compared to co-administration of free CpG ODN and antigen, we believe that this effect could be further improved. Here, we designed a novel delivery system of CpG ODN based on its conjugation to serve as anchor for liposomes. Thiol-maleimide chemistry was utilised to covalently ligate model protein with the CpG ODN TLR9 agonist. Due to its negative charge, the protein conjugate readily electrostatically bound cationic liposomes composed of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and dimethyldioctadecylammonium bromide (DDA) in a very high degree. The novel cationic liposomes-protein conjugate complex shared similar vesicle characteristics (size and charge) compared to free liposomes. The conjugation of CpG ODN to protein in conjunction with adsorption on cationic liposomes, could promote co-delivery leading to the induction of immune response at low antigen and CpG ODN doses. • The CpG ODN Toll-like receptor (TLR) 9 agonist was conjugated to protein antigens via thiol-maleimide chemistry. • Due to their negative charge, protein conjugates readily electrostatically bound cationic liposomes composed of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and dimethyldioctadecylammonium bromide (DDA) resulting to the design of novel cationic liposomes-protein conjugate complexes. • The method is suited for the liposomal delivery of a variety of adjuvant-protein conjugates. |
| first_indexed | 2024-12-14T14:32:48Z |
| format | Article |
| id | doaj.art-477d9d311e264ae7969c12e06318030c |
| institution | Directory Open Access Journal |
| issn | 2215-0161 |
| language | English |
| last_indexed | 2024-12-14T14:32:48Z |
| publishDate | 2020-01-01 |
| publisher | Elsevier |
| record_format | Article |
| series | MethodsX |
| spelling | doaj.art-477d9d311e264ae7969c12e06318030c2022-12-21T22:57:45ZengElsevierMethodsX2215-01612020-01-017100942Synthesis of protein conjugates adsorbed on cationic liposomes surfaceDespo Chatzikleanthous0Robert Cunliffe1Filippo Carboni2Maria Rosaria Romano3Derek T. O'Hagan4Craig W. Roberts5Yvonne Perrie6Roberto Adamo7Strathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral St, G4 0RE Glasgow, UKStrathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral St, G4 0RE Glasgow, UKGSK, Via Fiorentina 1, 53100 Siena, ItalyGSK, Via Fiorentina 1, 53100 Siena, ItalyGSK, 14200 Shady Grove Rd, Rockville, MD, United StatesStrathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral St, G4 0RE Glasgow, UKStrathclyde Institute of Pharmacy and Biomedical Sciences, University of Strathclyde, 161 Cathedral St, G4 0RE Glasgow, UKGSK, Via Fiorentina 1, 53100 Siena, Italy; Corresponding author.The well-known Toll like receptor 9 (TLR9) agonist CpG ODN has shown promising results as vaccine adjuvant in preclinical and clinical studies, however its in vivo stability and potential systemic toxicity remain a concern. In an effort to overcome these issues, different strategies have been explored including conjugation of CpG ODN with proteins or encapsulation/adsorption of CpG ODN into/onto liposomes. Although these methods have resulted in enhanced immunopotency compared to co-administration of free CpG ODN and antigen, we believe that this effect could be further improved. Here, we designed a novel delivery system of CpG ODN based on its conjugation to serve as anchor for liposomes. Thiol-maleimide chemistry was utilised to covalently ligate model protein with the CpG ODN TLR9 agonist. Due to its negative charge, the protein conjugate readily electrostatically bound cationic liposomes composed of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and dimethyldioctadecylammonium bromide (DDA) in a very high degree. The novel cationic liposomes-protein conjugate complex shared similar vesicle characteristics (size and charge) compared to free liposomes. The conjugation of CpG ODN to protein in conjunction with adsorption on cationic liposomes, could promote co-delivery leading to the induction of immune response at low antigen and CpG ODN doses. • The CpG ODN Toll-like receptor (TLR) 9 agonist was conjugated to protein antigens via thiol-maleimide chemistry. • Due to their negative charge, protein conjugates readily electrostatically bound cationic liposomes composed of 1,2-distearoyl-sn-glycero-3-phosphocholine (DSPC), cholesterol and dimethyldioctadecylammonium bromide (DDA) resulting to the design of novel cationic liposomes-protein conjugate complexes. • The method is suited for the liposomal delivery of a variety of adjuvant-protein conjugates.http://www.sciencedirect.com/science/article/pii/S221501612030162XTLR9 agonistConjugationCationic liposomesSurface adsorptionNanoparticlesVaccines |
| spellingShingle | Despo Chatzikleanthous Robert Cunliffe Filippo Carboni Maria Rosaria Romano Derek T. O'Hagan Craig W. Roberts Yvonne Perrie Roberto Adamo Synthesis of protein conjugates adsorbed on cationic liposomes surface MethodsX TLR9 agonist Conjugation Cationic liposomes Surface adsorption Nanoparticles Vaccines |
| title | Synthesis of protein conjugates adsorbed on cationic liposomes surface |
| title_full | Synthesis of protein conjugates adsorbed on cationic liposomes surface |
| title_fullStr | Synthesis of protein conjugates adsorbed on cationic liposomes surface |
| title_full_unstemmed | Synthesis of protein conjugates adsorbed on cationic liposomes surface |
| title_short | Synthesis of protein conjugates adsorbed on cationic liposomes surface |
| title_sort | synthesis of protein conjugates adsorbed on cationic liposomes surface |
| topic | TLR9 agonist Conjugation Cationic liposomes Surface adsorption Nanoparticles Vaccines |
| url | http://www.sciencedirect.com/science/article/pii/S221501612030162X |
| work_keys_str_mv | AT despochatzikleanthous synthesisofproteinconjugatesadsorbedoncationicliposomessurface AT robertcunliffe synthesisofproteinconjugatesadsorbedoncationicliposomessurface AT filippocarboni synthesisofproteinconjugatesadsorbedoncationicliposomessurface AT mariarosariaromano synthesisofproteinconjugatesadsorbedoncationicliposomessurface AT derektohagan synthesisofproteinconjugatesadsorbedoncationicliposomessurface AT craigwroberts synthesisofproteinconjugatesadsorbedoncationicliposomessurface AT yvonneperrie synthesisofproteinconjugatesadsorbedoncationicliposomessurface AT robertoadamo synthesisofproteinconjugatesadsorbedoncationicliposomessurface |