plate background becomes slightly yellowish. Carbohydrates
are stained in dark brown or black color.
11. Addition of sulfuric acid to the aqueous solution will generate
heat. Allow the sample to cool gradually to ambient temperature. Wait 20 min or until complete color change is seen before
transferring samples to a cuvette.
12. The addition of carrier protein in the blank aims to eliminate
the potential influence of protein on the carbohydrate analysis
of glycoconjugate.
13. This method can only provide qualitative results confirming
the conjugation.
References
1. Monzavi-Karbassi B, Pashov A, KieberEmmons T (2013) Tumor-associated glycans
and immune surveillance. Vaccine 1:174–203
2. Hakomori SI (2001) Tumor-associated carbohydrate antigens defining tumor malignancy:
basis for development of anti-cancer vaccines.
Adv Exp Med Biol 491:369–402
3. Danishefsky S, Allen J (2000) From the laboratory to the clinic: a retrospective on fully synthetic carbohydrate-based anticancer vaccines.
Angew Chem Int Ed Engl 39:836–863
4. Feng D, Shaikh AS, Wang F (2016) Recent
advance in tumor-associated carbohydrate antigens (TACAs)-based antitumor vaccines. ACS
Chem Biol 11:850–863
5. Guo Z, Wang Q (2009) Recent development
in carbohydrate-based cancer vaccines. Curr
Opin Chem Biol 13:608–617
6. Moyle PM, Toth I (2008) Self-adjuvanting
lipopeptide vaccines. Curr Med Chem
15:506–516
7. Hermanson GT (2013) The reactions of bioconjugation. In: Audet J, Preap M (eds) Bioconjugate techniques. Elsevier, Rockford, pp
229–258
8. Gamblin DP, Scanlan EM, Davis BG (2009)
Glycoprotein synthesis: an update. Chem Rev
109:131–163
9. Jacques S, Rich JR, Ling C et al (2006) Chemoenzymatic synthesis of GM 3 and GM
2 gangliosides containing a truncated ceramide
functionalized for glycoconjugate synthesis
and solid phase applications. Org Biomol
Chem 3:142–154
10. Andersen SM, Ling C-C, Zhang P et al (2004)
Synthesis of ganglioside epitopes for oligosaccharide specific immunoadsorption therapy of
Guillian-Barre ´ syndrome. Org Biomol Chem
2:1199–1212
11. Wang Q, Xue J, Guo Z (2009) Synthesis of a
monophosphoryl lipid A derivative and its conjugation to a modified form of a tumorassociated carbohydrate antigen GM3. Chem
Commun 5536–5537
12. Liao G, Zhou Z, Suryawanshi S et al (2016)
Fully
synthetic
self-adjuvanting
α-2,9-oligosialic acid based conjugate vaccines
against group C meningitis. ACS Cent Sci
2:210–218
13. Toyokuni T, Dean B, Cai S et al (1994) Synthetic vaccines: synthesis of a dimeric Tn
antigen-lipopeptide conjugate that elicits
immune responses against Tn-expressing glycoproteins. J Am Chem Soc 116:395–396
14. Wang Q, Ekanayaka SA, Wu J et al (2008)
Synthetic and immunological studies of 5
0 -Nphenylacetyl sTN to develop carbohydratebased cancer vaccines and to explore the
impacts of linkage between carbohydrate antigens and carrier proteins. Bioconjug Chem
19:2060–2067
15. Zhou Z, Liao G, Mandal SS et al (2015) A fully
synthetic self-adjuvanting globo H-based vaccine elicited strong T cell-mediated antitumor
immunity. Chem Sci 6:7112–7121
16. Liao G, Zhou Z, Burgula S et al (2015) Synthesis and immunological studies of linear oligosaccharides of β-glucan as antigens for
antifungal vaccine development. Bioconjug
Chem 26:466–476
17. Buskas T, Li Y, Boons G (2004) The immunogenicity of the tumor-associated antigen Lewis
(y) may be suppressed by a bifunctional crosslinker required for coupling to a carrier protein.
Chem Eur J 10:3517–3524
18. Burgula S, Swarts BM, Guo Z (2012) Total
synthesis of a glycosylphosphatidylinositol
Oligosaccharide-Protein Conjugation
311
are stained in dark brown or black color.
11. Addition of sulfuric acid to the aqueous solution will generate
heat. Allow the sample to cool gradually to ambient temperature. Wait 20 min or until complete color change is seen before
transferring samples to a cuvette.
12. The addition of carrier protein in the blank aims to eliminate
the potential influence of protein on the carbohydrate analysis
of glycoconjugate.
13. This method can only provide qualitative results confirming
the conjugation.
References
1. Monzavi-Karbassi B, Pashov A, KieberEmmons T (2013) Tumor-associated glycans
and immune surveillance. Vaccine 1:174–203
2. Hakomori SI (2001) Tumor-associated carbohydrate antigens defining tumor malignancy:
basis for development of anti-cancer vaccines.
Adv Exp Med Biol 491:369–402
3. Danishefsky S, Allen J (2000) From the laboratory to the clinic: a retrospective on fully synthetic carbohydrate-based anticancer vaccines.
Angew Chem Int Ed Engl 39:836–863
4. Feng D, Shaikh AS, Wang F (2016) Recent
advance in tumor-associated carbohydrate antigens (TACAs)-based antitumor vaccines. ACS
Chem Biol 11:850–863
5. Guo Z, Wang Q (2009) Recent development
in carbohydrate-based cancer vaccines. Curr
Opin Chem Biol 13:608–617
6. Moyle PM, Toth I (2008) Self-adjuvanting
lipopeptide vaccines. Curr Med Chem
15:506–516
7. Hermanson GT (2013) The reactions of bioconjugation. In: Audet J, Preap M (eds) Bioconjugate techniques. Elsevier, Rockford, pp
229–258
8. Gamblin DP, Scanlan EM, Davis BG (2009)
Glycoprotein synthesis: an update. Chem Rev
109:131–163
9. Jacques S, Rich JR, Ling C et al (2006) Chemoenzymatic synthesis of GM 3 and GM
2 gangliosides containing a truncated ceramide
functionalized for glycoconjugate synthesis
and solid phase applications. Org Biomol
Chem 3:142–154
10. Andersen SM, Ling C-C, Zhang P et al (2004)
Synthesis of ganglioside epitopes for oligosaccharide specific immunoadsorption therapy of
Guillian-Barre ´ syndrome. Org Biomol Chem
2:1199–1212
11. Wang Q, Xue J, Guo Z (2009) Synthesis of a
monophosphoryl lipid A derivative and its conjugation to a modified form of a tumorassociated carbohydrate antigen GM3. Chem
Commun 5536–5537
12. Liao G, Zhou Z, Suryawanshi S et al (2016)
Fully
synthetic
self-adjuvanting
α-2,9-oligosialic acid based conjugate vaccines
against group C meningitis. ACS Cent Sci
2:210–218
13. Toyokuni T, Dean B, Cai S et al (1994) Synthetic vaccines: synthesis of a dimeric Tn
antigen-lipopeptide conjugate that elicits
immune responses against Tn-expressing glycoproteins. J Am Chem Soc 116:395–396
14. Wang Q, Ekanayaka SA, Wu J et al (2008)
Synthetic and immunological studies of 5
0 -Nphenylacetyl sTN to develop carbohydratebased cancer vaccines and to explore the
impacts of linkage between carbohydrate antigens and carrier proteins. Bioconjug Chem
19:2060–2067
15. Zhou Z, Liao G, Mandal SS et al (2015) A fully
synthetic self-adjuvanting globo H-based vaccine elicited strong T cell-mediated antitumor
immunity. Chem Sci 6:7112–7121
16. Liao G, Zhou Z, Burgula S et al (2015) Synthesis and immunological studies of linear oligosaccharides of β-glucan as antigens for
antifungal vaccine development. Bioconjug
Chem 26:466–476
17. Buskas T, Li Y, Boons G (2004) The immunogenicity of the tumor-associated antigen Lewis
(y) may be suppressed by a bifunctional crosslinker required for coupling to a carrier protein.
Chem Eur J 10:3517–3524
18. Burgula S, Swarts BM, Guo Z (2012) Total
synthesis of a glycosylphosphatidylinositol
Oligosaccharide-Protein Conjugation
311