A First Case of Fluorescence Polarization Biosensor-Based Assay for Rapid Monitoring of Protein API Content in Tablet Dosage Forms: Detection of Lysozyme in Tablets

A First Case of Fluorescence Polarization Biosensor-Based Assay for Rapid Monitoring of Protein API Content in Tablet Dosage Forms: Detection of Lysozyme in Tablets Научная публикация

Журнал

Biosensors-Basel
ISSN: 2079-6374

Вых. Данные

Год: 2025, Том: 15, Номер: 11, Номер статьи : 724, Страниц : DOI: 10.3390/bios15110724

Авторы

Filimonova Svetlana M. ¹ , Balyklova Ksenia S. ¹ , Zherdev Dmitry O. ² , Eremin Sergei A. ² , Mukhametova Liliya I. ² , Krylov Vadim B. ³ , Nifantiev Nikolay E. ³

Организации

1	Institute of Pharmacy, Federal State Autonomus Educational Institution of Higher Education I.M. Sechenov First Moscow State Medical University of Ministry of Health of the Russian Federation (Sechenovskiy University), 119435 Moscow, Russia
2	Department of Chemistry, Lomonosov Moscow State University, 119991 Moscow, Russia
3	Laboratory of Glycoconjugate Chemistry, N.D. Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences, Leninsky Prospect 47, 119991 Moscow, Russia

Protein-based APIs represent a big group of modern therapeutics. Their characterization involves complex analytical protocols which require special methods, especially in the case when the protein drug is included into tablet dosage forms. Although the fluorescence polarization assay (FPA) is not currently regulated by many national Pharmacopeias, it represents a promising approach for protein drug standardization, considering their rapid, sensitive, and automatable detection suitable for high-throughput analysis and real-time quality control. To evaluate the applicability of FPA for the analysis of protein drugs in tablets, the quantifying of lysozyme in tablet dosage forms was studied by this method with the use of a fluorescently labeled synthetic chitooligosaccharide tracer. It was shown that this approach overcomes the limitations of the conventional turbidimetric assay of lysozyme determination, which is labor-intensive and relies on unstable reagents. Measurements were performed with both portable and stationary fluorescence polarization readers. Commercial tablets from five manufacturers containing lysozyme (20 mg) and pyridoxine hydrochloride (10 mg) together with other excipients were analyzed. The FPIA method showed a linear range of 5.0–70 µg/mL, with specificity confirmed by the absence of interference from excipients. Accuracy, evaluated by standard addition (10–20 mg), yielded recoveries of 100.2–106.0%. Placebo spiked with lysozyme at 80–120% of nominal content demonstrated recoveries of 98.0–100.1%, with RSD (n = 6) not exceeding 13.7%, indicating good precision. The developed method enables reliable lysozyme quantification in tablets, offering speed, simplicity, and robustness, and shows its suitability for the routine quality control of protein-containing dosage forms including the enzyme ones.

Filimonova S.M. , Balyklova K.S. , Zherdev D.O. , Eremin S.A. , Mukhametova L.I. , Krylov V.B. , Nifantiev N.E.
A First Case of Fluorescence Polarization Biosensor-Based Assay for Rapid Monitoring of Protein API Content in Tablet Dosage Forms: Detection of Lysozyme in Tablets
Biosensors-Basel. 2025. V.15. N11. 724 . DOI: 10.3390/bios15110724 WOS Scopus OpenAlex

≡ Web of science:	WOS:001624065700001
≡ Scopus:	2-s2.0-105022910806
≡ OpenAlex:	W4415822602