Home › Biochemistry › Comparative Analysis of Human Growth Hormone in Serum Using SPRi, Nano-SPRi and ELISA Assays
Biochemistry JoVE (Open Access) Citable · DOI

Comparative Analysis of Human Growth Hormone in Serum Using SPRi, Nano-SPRi and ELISA Assays

DOI: 10.3791/53508-v
What you'll learn
  • ✓ Compare SPRi, Nano-SPRi, and ELISA assay performance for hGH detection
  • ✓ Prepare and use SPRi chips with immobilized antibody arrays
  • ✓ Analyze recombinant human growth hormone in spiked serum samples
Protocol

The proposed work assesses the diagnostic potential of direct and amplified surface plasmon resonance imaging (SPRi) assays, particularly for the detection of recombinant human growth hormone in spiked human serum, by comparing SPRi results directly with commercially available enzyme-linked immunosorbent assay (ELISA) kit.

Difficulty
advanced
Total time
~4–6 hours per sample batch (chip preparation ~1 hr, SPRi detection ~1–2 hrs, ELISA ~2–3 hrs)
Biosafety
BSL-1

Steps

1
Prepare SPRi chip for antibody array

Clean and activate the SPRi sensor chip surface, then immobilize capture antibodies in an array format. This establishes the binding matrix required for direct analyte detection.

▶ 01:15
2
Detect human growth hormone via SPRi

Apply spiked human serum samples to the antibody-coated chip and measure real-time surface plasmon resonance imaging signal changes to quantify hGH binding.

▶ 04:08
3
Perform ELISA protocol for comparison

Execute a standard enzyme-linked immunosorbent assay using a commercial hGH kit on the same serum samples to generate a reference standard for assay validation.

▶ 05:46
4
Analyze and compare assay results

Evaluate SPRi, Nano-SPRi, and ELISA data in parallel, assessing sensitivity, specificity, and diagnostic agreement across the three detection platforms.

▶ 08:21

🚨 Failure Case Library (3) + Submit your own case

severe
B/B0 Ratio Outside Acceptable Range at Curve Endpoints
The B/B0 ratio (bound/maximum bound) for the terminal point of the standard curve is either >95% or <5%, indicating insufficient competitive displacement range.
💡 3 · ✓ 3
severe
Detection System Lacks Sensitivity for Target Concentration
Low or no signal despite proper assay setup. High concentration samples may show weak signal while low concentration samples are undetectable.
💡 4 · ✓ 4
minor
Sample Values Above Assay Range (Hook Effect Excluded)
Sample absorbance readings exceed the top standard, reading off the curve, while the standard curve itself appears normal with proper shape and dynamic range.
💡 2 · ✓ 3
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