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Vaccine. 2017 Mar 7;35(10):1410-1416. doi: 10.1016/j.vaccine.2017.01.077. Epub 2017 Feb 9.

A simple fluorescence-based assay for quantification of the Toll-Like Receptor agonist E6020 in vaccine formulations.

Author information

1
Departments of Pediatrics and Molecular Virology and Microbiology, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX 77030, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX 77030, USA.
2
Eisai Inc., RTP, Global Formulation Research, USA.
3
Departments of Pediatrics and Molecular Virology and Microbiology, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX 77030, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX 77030, USA; Department of Biology, Baylor University, Waco, TX, USA.
4
Departments of Pediatrics and Molecular Virology and Microbiology, National School of Tropical Medicine, Baylor College of Medicine, Houston, TX 77030, USA; Sabin Vaccine Institute and Texas Children's Hospital Center for Vaccine Development, Houston, TX 77030, USA; Department of Biology, Baylor University, Waco, TX, USA. Electronic address: bottazzi@bcm.edu.

Abstract

Despite the generally accepted immunostimulatory effect of Toll-Like Receptor 4 (TLR4) agonists and their value as vaccine adjuvants, there remains a demand for fast and easy quantification assays for these TLR4 agonists in order to accelerate and improve vaccine formulation studies. A new medium-throughput method was developed for the quantification of the TLR4 agonist, E6020, independent of the formulation composition. The assay uses a fluorescent hydrazide (DCCH) to label the synthetic lipopolysaccharide (LPS) analog E6020 through its diketone groups. This novel, low-cost, and fluorescence based assay may obviate the need for traditional approaches that primarily rely on Fourier transform infrared spectroscopy (FTIR) or mass spectrometry. The experiments were performed in a wide diversity of vaccine formulations containing E6020 to assess method robustness and accuracy. The assay was also expanded to evaluate the loading efficiency of E6020 in poly(lactic-co-glycolic acid) (PLGA) micro-particles.

KEYWORDS:

Adjuvant quantification; Fluorescent hydrazide labeling; TLR4 agonist

PMID:
28190745
DOI:
10.1016/j.vaccine.2017.01.077
[Indexed for MEDLINE]

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