Analysis of oral uptake of a synthetic adjuvant by an immunoassay with monoclonal antibodies.
A blocking ELISA with specific antibodies against a synthetic sulpholipo-polysucrose (SL-Ficoll400) was developed to quantify this vaccine adjuvant component in faeces with the purpose of estimating the uptake thereof by the gastrointestinal tract after oral administration. Specific monoclonal antibodies (MABs) prepared against SL-Ficoll400 did not cross-react with components in murine faeces. SL-Ficoll400 inhibited the reactivity of the MABs in a dose-dependent fashion in the blocking ELISA and the concentration causing 50% inhibition (IC50) was about 1 microg SL-Ficoll400 per milliliter. Derivatives of Ficoll400 lacking sulphate or lipid groups or both were unable to block the MABs (IC50 > 1000 microg/ml) which confirmed the specificity of the test system. Quantification of SL-Ficoll400 in extracts of faeces of untreated mice spiked with SL-Ficoll400 revealed that more than 90% could be recovered. The blocking ELISA was used to quantify SL-Ficoll400 in the faeces of mice given this compound orally. One, two and three days after oral administration, the cumulative recovery of SL-Ficoll400 in faeces was 30, 40 and 92%, respectively. From these data, we concluded that the MABs obtained were specific for SL-Ficoll400, that SL-Ficoll400 in murine faeces could be quantified by a blocking ELISA with MABs and that SL-Ficoll400 was poorly absorbed by the gastrointestinal tract.
Hilgers
L A
LA
Solvay Research and Technology, Applied Immunology, Brussels, Belgium. 1.a.t.hilgers@id.dlo.nl
Ghenne
L
L
Nicolas
I
I
Fochesato
M
M
Lejeune
G
G
Harmsen
M
M
Boon
B
B
D000276:Adjuvants, Immunologic / Q000493:pharmacokinetics* / Q000633:toxicity; D000284:Administration, Oral; D000818:Animals; D000911:Antibodies, Monoclonal; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005243:Feces / Q000737:chemistry; D005260:Female; D007408:Intestinal Absorption; D008070:Lipopolysaccharides / Q000276:immunology / Q000493:pharmacokinetics* / Q000633:toxicity; D051379:Mice
D000276:Adjuvants, Immunologic; D000911:Antibodies, Monoclonal; D008070:Lipopolysaccharides