PMID: 6973591Jan 1, 1981Paper

A radioactive antigen-binding assay for the measurement of antibody to Haemophilus influenzae type b capsular polysaccharide

Journal of Immunological Methods
J S KuoD W McCoy

Abstract

A new polyethylene glycol (PEG) radioimmunoprecipitation assay was developed for the detection of antibody to Haemophilus influenzae b capsular polysaccharide, polyribosylribitol phosphate (PRP). The radioactive antigen, [3H]PRP, with a high specific activity, was produced by growing the organism in the presence of [3H]ribose and was purified by hydroxylapatite and Sepharose 4B column chromatography. In the assay, PEG (12.5%) was used to separate antibody-bound [3H]PRP from free [3H]PRP. The assay covered the range of 0.5 and 20 ng antibody/assay at a maximum sensitivity of 0.5 approximately 1.0 ng antibody/assay. With various dilutions (1-20 ng antibody/assay) of S. Klein reference antiserum, the within-run coefficient of variation (CV) of 10 replicates ranged from 3.5 to 8.5%. Average CVs of 8.9% and 11.0% were obtained in the between-run and day-to-day reproducibility studies. The binding of [3H]PRP to S. Klein reference antiserum was severely inhibited by a minute amount of non-radioactive PRP; however, no significant interference was found in the presence of high concentrations of polysaccharides from Escherichia coli K100 and Streptococcus pneumoniae indicating that the RIA was highly specific for antibody to H. influenza...Continue Reading

References

Jan 1, 1977·Journal of Biological Standardization·K H WongE B Seligmann
Jan 1, 1979·Journal of Immunological Methods·S J WallerB S Andrews
Dec 1, 1968·The Biochemical Journal·D E Brundish, J Baddiley
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Citations

Jun 24, 1991·Journal of Immunological Methods·S RuthsT A Out
Dec 15, 2000·Vaccine·M J CaulfieldA Sabouraud
Dec 18, 1986·The New England Journal of Medicine·D M GranoffR S Daum
Aug 1, 1983·The Journal of Clinical Investigation·R S MunsonD M Granoff
Nov 30, 2007·Pediatrics International : Official Journal of the Japan Pediatric Society·Naruhiko IshiwadaYoichi Kohno
Feb 1, 1985·Journal of Clinical Microbiology·C A HimmelreichG A Storch
May 1, 1984·Infection and Immunity·N P SteeleR P Levine

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