Conjugation Chemistry

Lees, Andrew; Puvanesarajah, Velupillai; Frasch, Carl E.

doi:10.1128/9781555815820.ch11

Cited by 7 publications

(8 citation statements)

References 40 publications

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“…Prior to their conjugation, polysaccharides undergo size reduction by micro-fluidization in a high pressure homogenizer, which significantly reduces the viscosity and is likely to enhance the polysaccharides reactivity during conjugation224041. During the development of the quadrivalent conjugate vaccine used in this study, different conjugation chemistries have been used in coupling of the polysaccharides to the carrier protein using a spacer or direct linking them without a spacer.…”

Section: Methodsmentioning

confidence: 99%

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Solution conformation and flexibility of capsular polysaccharides from Neisseria meningitidis and glycoconjugates with the tetanus toxoid protein

Morris

Almutairi

Adams

et al. 2016

Sci Rep

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The structural integrity of meningococcal native, micro-fluidized and activated capsular polysaccharides and their glycoconjugates – in the form most relevant to their potential use as vaccines (dilute solution) - have been investigated with respect to their homogeneity, conformation and flexibility. Sedimentation velocity analysis showed that the polysaccharide size distributions were generally bimodal with some evidence for higher molar mass forms at higher concentration. Weight average molar masses Mw where lower for activated polysaccharides. Conjugation with tetanus toxoid protein however greatly increased the molar mass and polydispersity of the final conjugates. Glycoconjugates had an approximately unimodal log-normal but broad and large molar mass profiles, confirmed by sedimentation equilibrium “SEDFIT MSTAR” analysis. Conformation analysis using HYDFIT (which globally combines sedimentation and viscosity data), “Conformation Zoning” and Wales-van Holde approaches showed a high degree of flexibility – at least as great as the unconjugated polysaccharides, and very different from the tetanus toxoid (TT) protein used for the conjugation. As with the recently published finding for Hib-TT complexes, it is the carbohydrate component that dictates the solution behaviour of these glycoconjugates, although the lower intrinsic viscosities suggest some degree of compaction of the carbohydrate chains around the protein.

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Section: Methodsmentioning

confidence: 99%

“…The covalent binding of the polysaccharides and the spacer adipic acid dihydrazide (ADH) is carried out using a coupling chemistry by which the polysaccharides are activated under controlled conditions by a suitable cyanylating agent, 1-cyano-4-dimethylamino-pyridinium tetrafluoroborate (CDAP) see e.g 4142…”

Section: Methodsmentioning

confidence: 99%

Solution conformation and flexibility of capsular polysaccharides from Neisseria meningitidis and glycoconjugates with the tetanus toxoid protein

Morris

Almutairi

Adams

et al. 2016

Sci Rep

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“…For three important bacterial pathogens in particular, H. influenza type B (Hib), S. pneumoniae and N. meningitidis, glycoconjugates have proven to be highly effective in countries where they have been introduced (Ladhani 2012; Grijalva et al 2007). The standard production method for these conjugates involves the separate generation and purification of the protein and the carbohydrate moiety, chemical activation thereof, and conjugation as well as subsequent purification of the resulting glycoprotein (Lees et al 2008). Even though it is an established and accepted method, there are several drawbacks to this approach.…”

Section: Glycoprotein Expression In Bacterial Hosts: Current Applications and Future Opportunitiesmentioning

confidence: 99%

Bacterial Glycoengineering as a Biosynthetic Route to Customized Glycomolecules

Yates

Mills

DeLisa

2018

Advances in Biochemical Engineering/Biotechnology

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Bacteria have garnered increased interest in recent years as a platform for the biosynthesis of a variety of glycomolecules such as soluble oligosaccharides, surface-exposed carbohydrates, and glycoproteins. The ability to engineer commonly used laboratory species such as Escherichia coli to efficiently synthesize non-native sugar structures by recombinant expression of enzymes from various carbohydrate biosynthesis pathways has allowed for the facile generation of important products such as conjugate vaccines, glycosylated outer membrane vesicles, and a variety of other research reagents for studying and understanding the role of glycans in living systems. This chapter highlights some of the key discoveries and technologies for equipping bacteria with the requisite biosynthetic machinery to generate such products. As the bacterial glyco-toolbox continues to grow, these technologies are expected to expand the range of glycomolecules produced recombinantly in bacterial systems, thereby opening up this platform to an even larger number of applications.

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“…For example, thiol modified oligosaccharides can be readily conjugated to maleimide-containing CRM197 in phosphate buffer at room temperature to afford glycoconjugates. Alternatively, synthetic oligosaccharides containing a terminal amine are converted into the p -nitrophenyladipate ester 111 derivative and covalently coupled to, for example, lysine side chains in CRM197.…”

Section: Introductionmentioning

confidence: 99%

Discovery of Semi- and Fully-Synthetic Carbohydrate Vaccines Against Bacterial Infections Using a Medicinal Chemistry Approach

Seeberger

2021

Chem. Rev.

116

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The glycocalyx, a thick layer of carbohydrates, surrounds the cell wall of most bacterial and parasitic pathogens. Recognition of these unique glycans by the human immune system results in destruction of the invaders. To elicit a protective immune response, polysaccharides either isolated from the bacterial cell surface or conjugated with a carrier protein, for T-cell help, are administered. Conjugate vaccines based on isolated carbohydrates currently protect millions of people against Streptococcus pneumoniae , Haemophilus influenzae type b, and Neisseria meningitides infections. Active pharmaceutical ingredients (APIs) are increasingly discovered by medicinal chemistry and synthetic in origin, rather than isolated from natural sources. Converting vaccines from biologicals to pharmaceuticals requires a fundamental understanding of how the human immune system recognizes carbohydrates and could now be realized. To illustrate the chemistry-based approach to vaccine discovery, I summarize efforts focusing on synthetic glycan-based medicinal chemistry to understand the mammalian antiglycan immune response and define glycan epitopes for novel synthetic glycoconjugate vaccines against Streptococcus pneumoniae , Clostridium difficile , Klebsiella pneumoniae , and other bacteria. The chemical tools described here help us gain fundamental insights into how the human system recognizes carbohydrates and drive the discovery of carbohydrate vaccines.

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Conjugation Chemistry

Cited by 7 publications

References 40 publications

Solution conformation and flexibility of capsular polysaccharides from Neisseria meningitidis and glycoconjugates with the tetanus toxoid protein

Solution conformation and flexibility of capsular polysaccharides from Neisseria meningitidis and glycoconjugates with the tetanus toxoid protein

Bacterial Glycoengineering as a Biosynthetic Route to Customized Glycomolecules

Discovery of Semi- and Fully-Synthetic Carbohydrate Vaccines Against Bacterial Infections Using a Medicinal Chemistry Approach

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