[New protein assay with improved tolerability to interferences].

Area of Science:

• Biochemistry
• Analytical Chemistry

Background:

• Routine protein assays are often compromised by various interfering compounds.
• Existing protein quantification protocols require specific adjustments for different types of interference, complicating experimental procedures.

Purpose of the Study:

• To develop a simplified and robust protein assay method with improved tolerance to common interfering substances.
• To modify the classic Folin-Ciocalteu reagent for enhanced accuracy in protein concentration determination.

Main Methods:

• A new protein assay method was developed by modifying the components and concentrations of reagents based on the Folin-Ciocalteu reagent.
• Standard curves were generated using bovine serum albumin (BSA) to establish the method's performance.
• The method's tolerability to various interfering substances, including surfactants, chelators, reductants, and nitrogen-containing compounds, was systematically evaluated.

Main Results:

• The new method demonstrated accurate protein concentration detection even in the presence of high concentrations of common interfering substances.
• Specific examples of tolerated interferents include sodium dodecyl sulfate (SDS), NP-40, Triton X-100, EDTA, EGTA, Dithiothreitol (DTT), beta-Mercaptoethanol (ME), ammonium sulphate, and urea.
• The developed assay showed good stability and defined detectable wavelengths.

Conclusions:

• The novel protein assay method significantly enhances tolerance to a wide range of interfering substances.
• This improved robustness makes the method potentially valuable for accurate protein content measurement in complex biological samples previously challenging for standard assays.