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Molecular diagnostics using analytical immuno high performance liquid affinity chromatography

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Abstract

Analytical immuno high performance liquid affinity chromatography (analytical immuno HPLAC) was evaluated as a molecular diagnostic tool. Antibodies raised in rabbits against bovine neurophysin II were immobilized through Protein A crosslinking onto coated silica. Interaction of immobilized antibody with mobile antigen was characterized by zonal and frontal elutions of14C-labeled bovine neurophysin II under isocratic, nondenaturing conditions. The Chromatographic behavior shows that analytical immuno HPLAC with immobilized antibodies can be used to detect the number and functional nature of matrix-interacting antigens in mixtures, thus providing a quantitative Chromatographic technology for “antigen mapping”.

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Abbreviations

HPLAC:

high performance liquid affinity chromatography

IgG:

immunoglobulin G

BNPII:

bovine neurophysin II

HRP:

horseradish peroxidase

acim:

acetimidyl

PBS:

phosphate buffered saline

HPLC:

high performance liquid chromatography

WS-DCC:

water soluble cyclohexyl-3-(2-morpholino-ethyl) carbodiimide metho paratoluensulphonate

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Authors and Affiliations

  1. National Institute of Diabetes and Digestive and Kidney Diseases, National Institutes of Health, 20892, Bethesda, MD

    Paolo Caliceti, Giorgio Fassina & Irwin M. Chaiken

Authors
  1. Paolo Caliceti
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  2. Giorgio Fassina
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  3. Irwin M. Chaiken
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Caliceti, P., Fassina, G. & Chaiken, I.M. Molecular diagnostics using analytical immuno high performance liquid affinity chromatography. Appl Biochem Biotechnol 16, 119–128 (1987). https://doi.org/10.1007/BF02798361

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  • DOI: https://doi.org/10.1007/BF02798361

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