Characterizing Antibody Cross-reactivity for Immunoaffinity Purification of Analytes prior to Multiplexed Liquid Chromatography–Tandem Mass Spectrometry | Zendy

Thomas J. Laha | Zendy; Frederick G. Strathmann | Zendy; Zhican Wang | Zendy; Ian H. de Boer | Zendy; Kenneth E. Thummel | Zendy; Andrew N. Hoofnagle | Zendy

Open Access

Characterizing Antibody Cross-reactivity for Immunoaffinity Purification of Analytes prior to Multiplexed Liquid Chromatography–Tandem Mass Spectrometry

Author(s) -

Thomas J. Laha,

Frederick G. Strathmann,

Zhican Wang,

Ian H. de Boer,

Kenneth E. Thummel,

Andrew N. Hoofnagle

Publication year - 2012

Publication title -

clinical chemistry

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.705

H-Index - 218

eISSN - 1530-8561

pISSN - 0009-9147

DOI - 10.1373/clinchem.2012.185827

Subject(s) - chromatography , analyte , chemistry , mass spectrometry , tandem mass spectrometry , liquid chromatography–mass spectrometry , tandem , materials science , composite material

Immunoassays for 1α,25-dihydroxyvitamin D [1α,25(OH)(2)D] lack analytical specificity. We characterized the cross-reactivity of an anti-1α,25(OH)(2)D antibody with purified vitamin D metabolites and used these data to map the chemical features of 1α,25(OH)(2)D that are important for antibody binding. Additionally, we hypothesized that when combined with isotope-dilution liquid chromatography-tandem mass spectrometry (LC-MS/MS), antibody cross-reactivity could be used to semiselectively enrich for structurally similar metabolites of vitamin D in a multiplexed assay.

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