CYANAGEN Technology – Luminol-based Chemiluminescence

Luminol-based chemiluminescence is a technology that produces light via a chemical reaction between luminol and an oxidizing agent, and is commonly used to detect trace amounts of catalysts, such as iron in blood, or enzymes in labs. In a basic solution, luminol oxidizes, forming an excited-state product that emits blue light (approx. 425 nm) as it decays to its ground state.

Luminol-based chemiluminescence immunoassays offer significant advantages over traditional Radioimmunoassay (RIA), primarily due to superior safety, higher sensitivity, and faster processing times. While RIA was historically the gold standard, its reliance on radioactive isotopes created limitations that chemiluminescence has addressed.

Here are the key advantages of luminol chemiluminescence over RIA:

• No Radiation Safety Hazards: Unlike RIA, which requires handling hazardous, radioactive isotopes that pose health risks to laboratory staff, luminol chemiluminescence is safe, non-toxic, and does not require special, heavily regulated disposal procedures.
• Significantly Higher Sensitivity: Chemiluminescence can detect much lower concentrations of analytes, often reaching sensitivities of 10-18 mol/L, which is superior to RIA. This makes it ideal for detecting trace biologically active substances.
• Faster Results (Shorter Turnaround Time): The detection of the light signal in CLIA takes only a few seconds, allowing results to be completed within 9-60 minutes. RIA often requires long incubation periods, frequently taking several hours or days.
• Longer Reagent Shelf Life: Luminol reagents and conjugates are highly stable, often lasting for six months to over a year. Conversely, radioactive tracers in RIA have short half-lives, requiring frequent, expensive reordering.
• Wide Dynamic Range (Linearity): Chemiluminescence provides a broader linear range (up to 105 relative luminous units), meaning it can accurately measure very high and very low concentrations of analytes in a single sample without the need for extensive dilution.
• Higher Degree of Automation: CLIA systems are easily automated, allowing for high-throughput screening and increased workflow efficiency in modern laboratories.
• No Background Interference: Because the signal is produced by a chemical reaction without an external light source, there is minimal background interference, resulting in better signal-to-noise ratios.

Luminol Chemiluminescence Key Applications and Mechanisms
• Forensic Science (Blood Detection): Luminol is sprayed onto surfaces to detect latent bloodstains, where haemoglobin acts as a catalyst for the oxidation reaction.
• Biochemistry and Immunoassays: It is used in Western blotting and ELISA tests to detect proteins, DNA, or RNA, frequently using horseradish peroxidase (HRP) to catalyse the reaction for high-sensitivity detection.
• Environmental Monitoring: The technique acts as a biosensor to measure specific molecules.

Cyanagen developed three ECL product lines of substrates for Western blotting and chemiluminescent ELISA: WESTAR, WESTAR-ONE (both patented), and WESTAR-TAB. These non-isotopic, chemiluminescent substrates are designed for the chemiluminescent detection of immobilised proteins and nucleic acids conjugated to horseradish peroxidase (HRP).

The WESTAR (two-solution format) product line includes ETA C Ultra 2.0, the most sensitive ECL on the market with long signal duration up to 20 hours, and HYPERNOVA, the most potent signal intensity on the market up to ten times higher than all the competitors’ top-level kits.
The WESTAR-ONE (one-solution format) product line includes ONE-EXTREME, the most potent single-solution kit at the same level as competitors’ top-level two-solution ECLs.
The WESTAR-TAB (tablet format) is an innovative, unique product line of single-use ECL tablets.
The three product lines above are intended for research use only and shall not be used in any clinical procedures or diagnostic purposes prior to a dedicated development of a specific kit validated for diagnostics.

WESTAR (two-solution format), more details at http://bit.ly/4t5XREH
WESTAR-ONE (one-solution format), more details at https://bit.ly/3MdndzS
WESTAR-TAB (tablet format), more details at https://bit.ly/4qJy1ER

REQUESTS for samples: sales@cyanagen.com

References:

1. Kricka, L.J. (2000) Methods Enzymol. 305, 370-390.
2. Heindl, D. and Josel, H.P. (1997) Non-radioactive Analysis of Biomolecules, 258-261. Springer, Berlin.
3. Marzocchi, E., Grilli. S., Della Ciana, L., Prodi, L., Roda, A. and Mirasoli, M., (2008) Anal. Biochem., 377, 189-194.
4. Vdovenko, M. M., Della Ciana, L., Sakharov, I. Yu., (2009) Anal. Biochemistry, 392, 54-58.
5. Vdovenko, M. M., Della Ciana, L., Sakharov, I. Yu., (2010) Biotechnology Journal, 5(8),886-90.
6. Vdovenko, M.M., Zubkov, A.V., Kuznetsova, G.I., Della Ciana, L., Kuzmina, N.S., Sakharov, I. Yu., (2010) J Immunol Methods, 362 (1-2), 127-130.