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KMID : 0545120030130030469
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Journal of Microbiology and Biotechnology
2003 Volume.13 No. 3 p.469 ~ p.472
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Measurement of Free Polysaccharide in Tetanus Toxoid-Conjugate Vaccine Using Antibody/Ammonium Sulfate Precipitation
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Yoo TH
Kim HS/Park SS/Bang EY/Oh YK/Kim LS/Kim H/Hur BK/Ryu YW/Kim JS
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Abstract
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A method that effectively precipitates capsular polysaccharide of Haemophilus influenzae type b (polyribosylribitol phosphate PRP) conjugated to tetanus toxoid (TT) PRP TT in a liquid vaccine has been developed to measure free PRP present in TT-conjugate vaccine. The method involves adding anti-TT antibody and ammonium sulfate to precipitate PRP-TT conjugate and measuring free PRP in the supernatant. This new method provides a complete precipitation of the total PRP-TT and provides an accurate and reproducible measurements of free PRP. The accuracy of the assay was confirmed by spiking known amounts of unconjugated PRP to PRP-TT conjugate and the new method was found to have no effect on free PRP while precipitating PRP-TT. The published acid precipitation method did not produce reproducible results due to incomplete precipitation of PRP-TT especially when the vaccine is formulated in a salt-buffered solution.
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KEYWORD
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