Processing raw absorbance data generated by ELISA for measuring specific antibody activity

W. Steimer, B. Przyklenk, A. Bauernfeind

Research output: Contribution to journalArticlepeer-review

Abstract

The ELISA used was a system for the detection of IgG-antibodies against lipopolysaccharide (LPS) from Pseudomonas aeruginosa in patients suffering from cystic fibrosis. Rather than comparing absorbance values of unknown test samples to one or several reference samples, all samples were tested at several working dilutions previously distinguished as adequate. The absorbances of two dilutions were plotted against each other and linearized by an exponential function. Through repeated application of this function and following logit-log transformation, a linear scale was created with values proportional to antibody activity. The calculation is based on absorbances from the central part of the dose-response curve. Titres are multiples of the cut-off that was defined as titre 1, after surveying a control population, in which a log-normal distribution of absorbance values was observed. The method allows antibody quantitation over a range of up to five log10 dilutions in one assay.

Original languageEnglish
Pages (from-to)53-59
Number of pages7
JournalSerodiagnosis and Immunotherapy in Infectious Disease
Volume5
Issue number1
DOIs
StatePublished - Apr 1993

Keywords

  • ELISA
  • enzyme immunoassay, data processing, antibody activity, Pseudomonas aeruginosa, cystic fibrosis

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