| Autor: |
Seid, Robert, Boykins, Robert, Liu, Dai-Fang, Kimbrough, Kimberly, Hsieh, Chia-Lung, Eby, Ronald |
| Zdroj: |
Glycoconjugate Journal; December 1989, Vol. 6 Issue: 4 p489-497, 9p |
| Abstrakt: |
We have defined the nature of the covalent linkages in aHaemophilus influenzaetype b oligosaccharide-CRM197conjugate vaccine, designated HbOC. The conjugate was acid hydrolyzed to release a novel amino-acid derivative,Nε-(2-hydroxyethyl)lysine (OHEt-Lys), identifiable with an amino-acid analyzer. This amino-acid derivative was formed by reduction of Schiff bases formed betweenH. influenzaetype b oligosaccharides (HbO) and the lysyl ε-amino groups of CRM197(a non-toxic, cross-reactive variant of diphtheria toxin), followed by acid hydrolysis of HbOC. Quantification of OHEt-Lys per CRM197molecule allowed the determination of a covalency ratio, a useful parameter for evaluating the stoichiometry and consistency of HbOC preparations. Covalent association between HbO and CRM197was also demonstrated by the coincidence of immunoreactivity of gelelectrophoresed HbOC on a Western blot probed with anti-CRM197and anti-saccharide antisera. |
| Databáze: |
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