Related Experiment Videos
A luminescent bioassay for thyroid blocking antibodies.
N J Jordan1, C Rinderle, J Ashfield
1Department of Medical Biochemistry, UWCM and University Hospital of Wales NHS Trust, Cardiff, UK.
Clinical Endocrinology
|April 12, 2001
Summary
A new rapid luciferase-based bioassay using the lulu* cell line can detect thyroid blocking antibodies (TBAb), which are implicated in hypothyroidism. This method offers a faster alternative to current lengthy assays for clinical use.
Area of Science:
- Endocrinology
- Immunology
- Cell Biology
Background:
- Thyroid blocking antibodies (TBAb) contribute to hypothyroidism and transient neonatal hypothyroidism.
- Current methods for measuring TBAb are time-consuming and complex.
- A need exists for a more efficient and clinically applicable TBAb detection assay.
Purpose of the Study:
- To develop and validate a rapid, luminescence-based bioassay for detecting TBAb.
- To utilize a novel cell line, lulu*, for TBAb measurement.
- To assess the assay's suitability for clinical diagnostic services.
Main Methods:
- Transfection of Chinese hamster ovary (CHO) cells with human TSH-R and a luciferase reporter construct.
- Selection and characterization of the lulu* cell clone for optimal TSH response.
- Optimization and validation of the luminescent bioassay using patient sera and comparison with RIA-based methods.
Main Results:
- The lulu* cell line demonstrated a dose-dependent luciferase response to TSH stimulation.
- The assay successfully detected TBAb in 12 previously confirmed positive samples.
- Specificity was high, with low false positive rates in control sera (SLE, RA, MNG).
- Potential for false positives with TSAb-containing samples was identified, necessitating careful formula selection and potential dilution series analysis.
Conclusions:
- A novel lulu* cell line enables a rapid, luminescent bioassay for TBAb detection.
- The assay is suitable for clinical implementation, offering a faster alternative to existing methods.
- Discrepancies in TBAb prevalence reported in literature may stem from variations in assay methodologies and calculation formulas.