Abstract:
:Factors influencing the performance of a dipstick competitive particle capture immunoassay (PCI) for the steroid oestrone sulphate (OS) were investigated. Appropriate 'blocking' of the nitrocellulose dipstick membrane was necessary for the upward flow of microsphere particles. Traditional protein blocking agents including BSA, gelatin and casein were unsatisfactory while synthetic polymers and surfactants were effective in promoting microsphere movement. A simple buffer consisting of 1% aqueous NaCl containing 0.05% Tween 20 was suitable for carrying the components up the dipstick and facilitating the antibody-antigen interactions. Increasing microsphere diameters from 0.3 to 0.8 microm allowed the microsphere antibody coating concentration to be reduced which enabled lower concentrations of OS to be measured. However, upward flow rate and the maximum signal attainable was compromised as a consequence. Enlarging the dipstick membrane nominal pore size from 3 to 12 microm increased the speed of test dot development, but assay sensitivity suffered as a result in some instances. Changing the capture antigen markedly influenced the dose-response lines. No dose-response was achieved with OG-BSA as the capture antigen while OHS-BSA and OCMO-BSA as capture antigens produced dose-response lines with means +/- S.E.M. EC(50) values of 140 +/- 16 and 19 +/- 1 ng/ml, respectively.
journal_name
J Immunol Methodsjournal_title
Journal of immunological methodsauthors
Henderson K,Stewart Jdoi
10.1016/s0022-1759(02)00280-6keywords:
subject
Has Abstractpub_date
2002-12-01 00:00:00pages
77-84issue
1eissn
0022-1759issn
1872-7905pii
S0022175902002806journal_volume
270pub_type
杂志文章abstract::We developed a dual wavelength spectrophotometric assay that permitted beta-lactamase and beta-galactosidase activities to be measured concurrently in a single sample. We also constructed a target cell, E. coli ML-35p, that was substantially cryptic for its periplasmic beta-lactamase and cytoplasmic beta-galactosidase...
journal_title:Journal of immunological methods
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journal_title:Journal of immunological methods
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