ISSN:
1573-4986
Keywords:
GM3 antigen
;
sialyllactoside
;
biantennary
;
glycoconjugate
;
antibody
;
CTP, Cytidine 5′-triphosphate
;
KLH, Keyhole Limpet Hemocyanin
;
M2C2H, 4-(4-N-maleimidomethyl) cyclohexane-1-carboxyl hydrazide
;
MPL, Monophosphoryl lipid A
;
Sulfo-GMBS, N-(γ-maleimidobutyryloxy) sulfosuccinimide ester
;
BSA, Bovine serum albumin
;
HSA, Human serum albumin
;
GBSPIa (GBSPIII), Type Ia (III) group B Streptococcus polysaccharide
Source:
Springer Online Journal Archives 1860-2000
Topics:
Chemistry and Pharmacology
Notes:
Abstract A biantennary GM3-saccharide (sialyllactoside) derivative (4) was constructed using allylmalonic acid as a bivalent linker, both carboxylic acids of which were condensed with 3-aminopropyl lactoside (2) prior to enzymatic sialylation with a fusion enzyme. While ozonolysis of its allyl group generated a saccharide having a terminal aldehyde (6), we were unable to couple 6 directly to protein by reductive amination. However, extension of the spacer by means of introducing a maleimide group to 6 through its aldehyde group to give 7 enabled the latter to be successfully coupled to thiolated proteins. The average ratios of saccharide to protein were observed to be 35 in KLH conjugate (13) and 9–12 in HSA conjugates (14 and 15). The antisera obtained by immunizing mice with the biantennary sialyllactoside-KLH conjugate (13) together with MPL adjuvant were analyzed by ELISA. Using several structurally related saccharide-HSA conjugates as screening antigens, it was concluded that anti-sialyllactoside antibodies, both IgG and IgM, were effectively raised. This was further supported by competitive inhibition experiments using lactoside (1), sialyllactoside (8) and biantennary sialyllactoside (4) as inhibitors.
Type of Medium:
Electronic Resource
URL:
http://dx.doi.org/10.1023/A:1007069904462
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