Where to find a sensitive Hyaluronic Acid (HA) quantitative assay

Hyaluronic acid (also called HA or Hyaluronan) is a glycosaminoglycan with elevated viscosity, enabling tissues (eye, skin, joint and synovial fluid…) to resist to physical and mechanical constraints (torsion, flexion…). Over time, when HA production declines, tissues progressively lose these tensile properties, leading to wrinkles and fold, weak re-epithelisation and age-related troubles. But HA is also involved in many other chronic and cancer-related diseases. In this post, we’ll review one of the most popular HA quantification assays (ELISA test), known to be highly sensitive and robust, appreciated by researchers involved in cosmetology and drug discovery.

Hyaluronan molecules are polymers of disaccharides (up to 25,000 disaccharide repeat units in length). These polymers have numerous applications in healthcare.  HA is one of several glycosaminoglycan components of the extracellular matrix of connective tissue.

Hyaluronic acid (HA) – a biomarker to follow in cosmetology & in Drug discovery

Skin cross section

Skin care and cosmetology R&D programs. Source: tebu-bio.

Known to be an anti-ageing biomarker in cosmetology, numerous studies have shown that HA is also involved in many chronic liver diseases, including infection (hepatitis B or C), toxicity (alcohol and drugs), genetic (hemochromatosis), autoimmunity and malignancy, liver fibrosis and cirrhosis. Impairment of liver function often results in a rapid increase in circulating HA levels. Additional data also indicates a relationship between HA levels, local inflammation and severity of diseases (e.g. HA levels in urine are indicative of bladder cancer, or can also be directly correlated to liver disease or during rheumatoid arthritis).

This makes HA a very popular target / biomarker in Life Sciences, for example:

  • Pharmaceutical approaches: Hyaluronan has been described in orthopaedics, eye surgery, wound healing, and regenerative medicine, chronic liver diseases and ageing-associated diseases…
  • Cosmetology and skin-care: HA is considered as an excellent moisturiser with anti-wrinkle properties

HA (Hyaluronan) measurement in biological samples is a meaningful biological marker (biomarker) for researchers involved in these research domains. But what are the criteria to perform reliable HA quantification in R&D programs?

Criteria for selecting your HA quantitative tests

#1 – Prefer immunoassays

Colorimetric protein Assay plate

ELISA tests to quantify Hyaluronic acid and many other biomarkers (Collagen, BDNF, CT-1…).

The most efficient methods to quantify HA still rely on antibody-based assays (e.g. Competitive or Sandwich ELISA test). Very convenient, these assays have many benefits – they come ready-to-use, integrate all the reagents needed to perform the quantification and are fully calibrated (HA for standard curve is included in the kits).

The golden 96-well format Hyaluronan Enzyme-Linked Immunosorbent Assay (cat. nr. 117K-1200-1ea) offers a simple yet very effective method for determining HA levels in human and animal biological fluids or cell supernatants.

This HA competitive ELISA test works best with HA molecules that are greater than 25 repeating units with a sensitivity of 25 ng/ml.

#2- Consider the volume of the sample

Classically, 100µl are required for running HA ELISA (meaning 200µl of sample if you work in duplicate). If your samples are smaller in size, you might consider the Sandwich Hyaluronan ELISA format from Echelon (cat. nr. K-4800-1kit).
This HA ELISA is a quantitative immunoassay especially designed for small volume (25 µL of sample per duplicate) or for the detection of 10+ disaccharide repeat units of HA. The lower detection limit is 25 ng/ml (Intra Coefficient of Variation  <5%  & Inter Coefficient of Variation <20%).
This makes this HA sandwich ELISA test ideal for in vitro measurements in conditions where HA is newly synthesized (cell cultures or human/animal biological fluids…) or when sample volumes are small.

#3- Outsource to skilled lab teams

More and more, research labs outsource their biomarker quantification to Contract Research Organizations (CRO). This choice is a clever and affordable solution when internal resources or competencies are lacking or when an overload of work is “rocking” a research program.

You might indeed prefer to outsource your HA quantification and send your samples to a service provider close to your laboratories.

Make sure that this outsourcing partner has a real presence (avoid online platforms promising very low costs and yet claiming good and rapid results…). Make sure that the company you choose does not further outsource themselves, or ship samples to other distant places. For example, tebu-bio laboratories are located in Europe (Paris area), their lab staff is used to working under Good Laboratory Practices (GLP) and ISO 9001:2008 conditions. They perform HA assays (but also assays for other markers like Collagen, Oxidative markers, MMPs, ORAC, ABTS, PPAR, ECM…) on a regular basis.

Learn more about their in vitro ingredient testing in cosmetology.

 

Want to reliably quantify Hyaluronic acid (HA) – Hyaluronan?

So, you can quantify HA levels in your samples yourself in your lab, or outsource it to tebu-bio’s lab staff who are experienced in running ELISA test and profiling assays.

Whatever your choice, it’s now as easy as ABC!

Philippe Fixe, PhD
Written by Philippe Fixe, PhD
Philippe Fixe is Marketing Director at tebu-bio, passionate about innovation and R&D in Life sciences, Biotechnology, Medical research, Drug discovery, and also a keen photographer.