Analysis of a monophosphoryl lipid A immunostimulant preparation from Salmonella minnesota R595 by high-performance liquid chromatography

Journal of Chromatography. a
S R HagenK R Myers

Abstract

MPL immunostimulant, a 4'-monophosphoryl lipid A (MLA) preparation obtained from the lipopolysaccharide of Salmonella minnesota R595, is being developed for several clinical indications. MPL comprises a mixture of MLA congeners that contain 4, 5, and 6 fatty acids. In this paper, we report a new high-performance liquid chromatography (HPLC) method for analyzing the congener composition and purity of MPL. MPL is first derivatized with dinitrobenzyloxyamine (DNBA), resulting in incorporation of the dinitrobenzyl chromophore at the reducing end of all MLA congeners. DNBA-MPL is then analyzed by reversed-phase HPLC on a Waters NovaPak C18, 4 microns particle size, 300 mm x 3.9 mm column. Optimal separation of DNBA-MLA species is obtained using a linear gradient of 10% to 80% isopropanol-water (95:5, v/v), 5 mM tetrabutylammonium dihydrogenphosphate (TBAP), in acetonitrile-water (95:5, v/v), 5 mM.TBAP, over 45 min. A synthetic compound, corresponding to a hexaacyl MLA congener, is used for determination of the detector response factor, allowing the MLA content of MPL (i.e., purity) to be determined. Overall, this method provides better separation, higher sensitivity, and is faster and saler than previous methods used for the analysi...Continue Reading

References

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Citations

Jul 26, 2003·Journal of Chromatography. B, Analytical Technologies in the Biomedical and Life Sciences·Yonghui WangJohn D Orr
Mar 23, 2006·Infection and Immunity·Liana TsenovaGilla Kaplan
May 3, 2013·PloS One·Carolyn R Casella, Thomas C Mitchell
Feb 20, 2014·PloS One·Christopher H CleggSteven G Reed
Jan 18, 2005·Journal of Controlled Release : Official Journal of the Controlled Release Society·Carrie S W ChongJohn Samuel
Feb 13, 2019·APMIS : Acta Pathologica, Microbiologica, Et Immunologica Scandinavica·Nastaran TaleghaniHoma Zamani
Jul 28, 2013·Expert Review of Vaccines·Christopher H CleggSusan L Baldwin
Jun 10, 2004·The Journal of Immunology : Official Journal of the American Association of Immunologists·Yasir A W SkeikySteven G Reed
Nov 13, 2020·Frontiers in Immunology·Yi-Qi WangThomas C Mitchell
Dec 2, 2019·Metabolic Engineering·Yuhyun JiHak Suk Chung
May 11, 2021·The Journal of Allergy and Clinical Immunology. in Practice·Oliver PfaarStefan Schülke
Dec 2, 1999·Journal of Medicinal Chemistry·D A JohnsonV G Brookshire

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