Synthesis ofStreptococcus pneumoniae Type 3 Neoglycoproteins Varying in Oligosaccharide Chain Length, Loading and Carrier Protein

Lefeber, Dirk; Kamerling, Johannis P.; Vliegenthart, Johannes F.G.

doi:10.1002/1521-3765(20011015)7:20<4411::aid-chem4411>3.0.co;2-t

Cited by 53 publications

(46 citation statements)

References 38 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…in their pioneering work . There are, however, examples in the literature of conjugations at higher pH . To examine the effect of pH on the efficiency of conjugation, we performed parallel conjugations with two different carrier proteins, BSA and rTT‐Hc at pH 8.5, 9.0, and 9.5, while targeting a molar carbohydrate/protein ratio (i.e., loading) of 5:1.…”

Section: Resultsmentioning

confidence: 99%

Conjugate Vaccines from Bacterial Antigens by Squaric Acid Chemistry: A Closer Look

Kelly

Vann³

et al. 2017

ChemBioChem

View full text Add to dashboard Cite

Using O-SP–core (O-SPcNH2) polysaccharide, isolated from Vibrio cholerae O1 lipopolysaccharide (LPS) and related synthetic substances, a detailed study of factors that affect conjugation of bacterial polysaccharides to protein carriers by squaric acid chemistry to form conjugate vaccines has been carried out. Several processes previously unrecognized, which take place during the squarate labelling of the O-SPcNH2 and subsequent conjugation of the squarate (O-SPcNH–SqOMe) formed, have been identified. The efficiency of conjugation at pH 8.5, 9.0, and 9.5 to each of bovine serum albumin (BSA) and the recombinant tetanus toxin fragment C (rTT-Hc) has been determined. The study led to a protocol for more efficient labeling of O-SPcNH2 antigen with the methyl squarate group, to yield a higher quality, more potent squarate conjugation reagent. Its use resulted in about two-fold increase of conjugation efficiency (from 23–26% on BSA to 51% on BSA and 55% on rTT-Hc). The spent conjugation reagent could be recovered and regenerated by treatment with MeI in the absence of additional base. The immunological properties of the experimental vaccine made from the regenerated conjugation reagent were comparable to those of the immunogen made from the parent O-SPcNH–SqOMe.

show abstract

Section: Resultsmentioning

confidence: 99%

Conjugate Vaccines from Bacterial Antigens by Squaric Acid Chemistry: A Closer Look

Kelly

Vann³

et al. 2017

ChemBioChem

View full text Add to dashboard Cite

show abstract

“…The glycosylation between 21 and 23 , [51] the precursor of the GlcA unit, went well to provide disaccharide 24 in high yield. The new benzylation reaction was performed on disaccharide 24 to readily provide the desired tetra-benzylated disaccharide 25 in 72% yield.…”

Section: Resultsmentioning

confidence: 99%

Regioselective Benzylation of 2‐Deoxy‐2‐aminosugars using Crown Ethers: Application to a Shortened Synthesis of Hyaluronic Acid Oligomers

2014

View full text Add to dashboard Cite

The combination of benzyl bromide, sodium hydroxide and 15-crown-5 in tetrahydrofuran is shown to be an efficient method for installing benzyl groups at both the 4- and 6-positions regioselectively directly from peracetylated N-trichloroacetyl-protected glucosamine and galactosamine. Application of this benzylation strategy proved to significantly shorten the synthetic route to hyaluronic acid tetra- and hexamers.

show abstract

“…The glycan epitopes ST3, PS1 and LPG ( Table 1 ) were synthesised from monosaccharide building blocks as described previously 43 44 45 .…”

Section: Methodsmentioning

confidence: 99%

Cross Reactive Material 197 glycoconjugate vaccines contain privileged conjugation sites

Möginger

Resemann

Martin

et al. 2016

Sci Rep

View full text Add to dashboard Cite

Production of glycoconjugate vaccines involves the chemical conjugation of glycans to an immunogenic carrier protein such as Cross-Reactive-Material-197 (CRM197). Instead of using glycans from natural sources recent vaccine development has been focusing on the use of synthetically defined minimal epitopes. While the glycan is structurally defined, the attachment sites on the protein are not. Fully characterized conjugates and batch-to-batch comparisons are the key to eventually create completely defined conjugates. A variety of glycoconjugates consisting of CRM197 and synthetic oligosaccharide epitopes was characterised using mass spectrometry techniques. The primary structure was assessed by combining intact protein MALDI-TOF-MS, LC-MALDI-TOF-MS middle-down and LC-ESI-MS bottom-up approaches. The middle-down approach on CNBr cleaved glycopeptides provided almost complete sequence coverage, facilitating rapid batch-to-batch comparisons, resolving glycan loading and identification of side products. Regions close to the N- and C-termini were most efficiently conjugated.

show abstract

Synthesis ofStreptococcus pneumoniae Type 3 Neoglycoproteins Varying in Oligosaccharide Chain Length, Loading and Carrier Protein

Cited by 53 publications

References 38 publications

Conjugate Vaccines from Bacterial Antigens by Squaric Acid Chemistry: A Closer Look

Conjugate Vaccines from Bacterial Antigens by Squaric Acid Chemistry: A Closer Look

Regioselective Benzylation of 2‐Deoxy‐2‐aminosugars using Crown Ethers: Application to a Shortened Synthesis of Hyaluronic Acid Oligomers

Cross Reactive Material 197 glycoconjugate vaccines contain privileged conjugation sites

Contact Info

Product

Resources

About