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Updated: Aug 8, 2026
Comparative Analysis of Human Growth Hormone in Serum Using SPRi, Nano-SPRi and ELISA Assays
Published on: January 7, 2016
Time-resolved immunofluorometric assay of human growth hormone
K Albertsson-Wikland1, C Jansson, S Rosberg
1Department of Pediatrics, University of Göteborg, Sweden.
Insights
A new time-resolved immunofluorometric assay (trIFMA) for human growth hormone (hGH) offers improved sensitivity and speed. This assay uses monoclonal antibodies (mAbs) and is ideal for pediatric use due to its small serum volume requirement.
Area of Science:
- Biochemistry
- Immunology
- Analytical Chemistry
Background:
- Accurate measurement of human growth hormone (hGH) is crucial for diagnosing endocrine disorders in children.
- Existing immunoradiometric assays (IRMA) have limitations in sensitivity and require larger sample volumes.
- Development of a more sensitive and efficient assay for hGH is needed, particularly for pediatric applications.
Purpose of the Study:
- To develop and validate a time-resolved immunofluorometric assay (trIFMA) for quantifying human growth hormone (hGH).
- To compare the performance of the novel trIFMA with established immunoradiometric assays (IRMA).
- To assess the suitability of the trIFMA for pediatric use, considering sensitivity and sample volume requirements.
Main Methods:
- A time-resolved immunofluorometric assay (trIFMA) was developed using monoclonal antibody (mAb)-coated microtiter strip wells and a europium (Eu) chelate-labeled mAb.
- The trIFMA was compared to an immunoradiometric assay (IRMA) utilizing polyclonal antibodies.
- Serum samples from 36 children with various diagnoses (n=185) and 24-h profile samples from 39 children were analyzed.
Main Results:
- The trIFMA demonstrated higher sensitivity with a detection limit of 0.03 mIU/L, surpassing existing IRMAs.
- Assay precision was good, with intra- and interassay coefficients of variation (CVs) below 10.6% for hGH concentrations between 1 and 100 mIU/L.
- The trIFMA is rapid (1s counting time), technically simple, requires no centrifugation, and uses only 25 μL of serum, making it suitable for pediatric use.
Conclusions:
- The developed trIFMA is a sensitive, rapid, and technically simple assay for hGH measurement.
- Its high sensitivity, low serum volume requirement, and ease of use make it particularly advantageous for pediatric endocrinology.
- The assay selectively detects the 22-kDa form of hGH, accounting for approximately 80% of the amount measured by polyclonal IRMA.
Abstract:
We describe a time-resolved immunofluorometric assay (trIFMA) for human growth hormone (hGH), in which monoclonal antibody (mAb)-coated microtiter strip wells and a europium (Eu) chelate-labeled mAb are used. We compare the new trIFMA, in which two mAbs are used, with an immunoradiometric assay (IRMA) in which polyclonal antibodies are used. Serum samples (n = 185) from 36 children with various diagnoses were analyzed. In addition, 24-h profile samples (72 per child) from 39 children were analyzed. The trIFMA was more sensitive (detection limit, 0.03 mIU/L) than existing IRMAs. Both the intra- and interassay CVs were < or = 10.6% for hGH concentrations between 1 and 100 mIU/L. The trIFMA is technically simple and rapid, requires no centrifugation or separation reagent, and has a counting time of only 1 s per sample. In addition, the Eu label is nontoxic, presents no waste-disposal problems, and has a long shelf-life. Finally, the assay requires only small volumes of serum (25 muL), which is of considerable importance in pediatric use. The mAbs used for the trIFMA selectively bind the 22-kDa form of hGH, with the result that the assay detects about 80% of the amount detected by the polyclonal IRMA.
