Archives of Toxicology

, Volume 64, Issue 5, pp 360–364 | Cite as

Immunoassays for proteins alkylated by nicotine-derived N-nitrosamines

  • Brian Talbot
  • Serge Desnoyers
  • Andre Castonguay
Original Investigations

Abstract

Polyclonal antibodies recognizing the pyridyloxobutyl (POB) moiety of 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK) were produced in rabbits immunized either with POB-bovine albumin or POB-Sepharose. The POB intermediates necessary to modify the protein were generated by alkaline (pH 9.0) treatment of the synthetic precursor 4-(carbethoxynitrosamino)-1-(3-pyridyl)-1-butanone. In a competitive enzyme linked immunoabsorbent assay (ELISA), 70 pmole NNK inhibited 50% of the binding of the anti-POB antibodies to POB-protein absorbed on microtiterplates. This 50% inhibition varied from 70 pmole to 200 nmole using a series of NNK analogues, depending on the integrity of the POB moiety. Immunological techniques initiated in this study detect NNK-protein conjugates or measure the quantity of POB groups liberated upon alkaline or acid treatment of NNK modified protein.

Key words

Carcinogen-protein adducts Tobacco-specific N-nitrosamines Dosimeter of carcinogen exposure Immunoassay 

Abbreviations

NNN

N′-nitrosonomicotine

NNK

4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone

NNK-N-oxide

4-(methylnitrosamino)-1-(3-pyridyl-N-oxide)-1-butanone

NNAL

4-(methylnitrosamino)-1-(3-pyridyl) butan-1-ol

ELISA

Enzyme linked immunosorbent assay

SDS

Sodium dodecyl sulfate

POB

pyridyloxobutyl

PBS

phosphate buffered saline

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Copyright information

© Springer-Verlag 1990

Authors and Affiliations

  • Brian Talbot
    • 1
  • Serge Desnoyers
    • 1
  • Andre Castonguay
    • 2
  1. 1.Department of Biology, Faculty of SciencesUniversity of SherbrookeSherbrookeCanada
  2. 2.Laboratory of Cancer Etiology and Chemoprevention, School of PharmacyLaval UniversityQuebec Q. CCanada

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