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Characterization of specific lysophosphatidic acid subspecies produced by autotaxin using a novel LC/MS/MS method
Author(s) -
Mayton Eric,
Wijesinghe Dayanjan,
Mietla Jennifer,
Fang Xianjun,
Chalfant Charles
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.541.2
Lysophosphatidic acid is a signaling molecule that has a plethora of biological functions including roles in cell survival, proliferation, and migration. LPA has been found to be dysregulated in a variety of malignancies such as cancer, atherosclerosis, and many more. Although HPLC/MS/MS technology has been used to measure the levels of LPA in human blood serum and plasma, there have been limited methods to detect minute levels of LPA from cell culture, restricting the ability of researchers to investigate specific metabolic and signaling pathways attributed to particular LPA subspecies. In this study, we developed a novel HPLC/MS/MS method with enhanced sensitivity that allows accurate measurements of LPA levels with a limit of detection for non‐naturally occurring standards between 0.1–1 attomoles. The method was highly sensitive with a limit of quantitation at approximately 1 attomole. The method was validated by examination of previously characterized cell lines ectopically expressing autotaxin which showed a three‐fold increase in femtomolar LPA levels. This new technology will allow researchers to measure in vitro LPA levels and also allow researchers to distinguish specific LPA subspecies.