Research paper
Monoclonal antibody development for acrylamide-adducted human
haemoglobin; A biomarker of dietary acrylamide exposure
Andrew Preston
a,
⁎
, Terry Fodey
b
, Alistair Douglas
b
, Christopher T. Elliott
c
a
Department of Biomedical Sciences, University of Liverpool, Sherrington Buildings, Ashton Street, Liverpool L69 3GE, UK
b
Agri-Food and Biosciences Institute, Stoney Road, Belfast BT4 3SD, UK
c
Institute of AgriFood and Land Use, Queen's University Belfast, Belfast BT7 1NN, UK
article info abstract
Article history:
Received 9 June 2008
Received in revised form 9 October 2008
Accepted 14 October 2008
Available online 18 November 2008
The spontaneous formation of the neurotoxic carcinogen acrylamide in a wide range of cooked
foods has recently been discovered, leading to dietary exposure estimates of 30.8 µg of
acrylamide day
− 1
for an average 77 kg human male. This is considerably higher than the
European legal limit of acrylamide in drinking water, which is approximately 0.2 µg of
acrylamide person
− 1
day
− 1
. A recent study of 62,573 women over 11.3 years has observed an
increased risk of postmenopausal endometrial and ovarian cancer (but not breast cancer) with
increasing dietary acrylamide intake, demonstrating significant risk to human health. As
individual acrylamide exposure is affected by dietary habits, cooking methods, and cigarette
consumption; accurate extrapolation from estimated dietary exposure is extremely difficult.
Quantifying biomarkers of acrylamide exposure therefore remains the most effective means of
rapidly determining individual exposure to acrylamide, and correlating exposure with lifestyle
choices. Current methodologies for the analysis of blood biomarkers of acrylamide are focused
on expensive, slower chromatographic techniques such as GC and LC coupled to mass
spectrometry. This paper describes the first successful development of two monoclonal
antibodies specific to acrylamide-adducted haemoglobin (IC
50
of 94 ng ml
− 1
and 198 ng ml
− 1
),
that are suitable for use in a high-throughput biomarker immunoassay to determine individual
acrylamide exposure. Further development of acrylamide-haemoglobin standards with defined
levels of acrylamide adduction will enable a fully quantitative assay, and allow sensitivity
comparisons with alternative chromatographic methods of analysis.
© 2008 Elsevier B.V. All rights reserved.
Keywords:
Acrylamide
Haemoglobin adduct
mAb
Biomarker
1. Introduction
The spontaneous formation of acrylamide (a neurotoxic
carcinogen) during the cooking of food has led to its description
as a cooking carcinogen (Tareke et al., 2000). Its widespread
presence has been uncovered in a range of processed foods
(SNFA, 2002; Zhang et al., 2005; Young et al., 2007), leading to
dietary exposure estimates of 0.4 µg kg
−1
of bodyweight day
−1
(Dybing et al., 2005), equivalent to 30.8 µg of acrylamide day
−1
for an average 77 kg human male. This is considerably higher
than the European legal limit of acrylamide in drinking water,
which is 0.1 µg L
−1
(EC Council Directive, 1998), or approxi-
mately 0.2 µg of acrylamide person
−1
day
−1
.Inadditiontothe
evidence of acrylamide's mutagenicity (Knaap et al., 1988;
Besaratinia and Pfeifer, 2003), genotoxicity (Dearfield et al.,
1988), and carcinogenicity (Segerbäck et al., 1995) in animals, a
recent study of 62,573 women over 11.3 years has observed an
increased risk of postmenopausal endometrial and ovarian
cancer (but not breast cancer) with increasing dietary acryla-
mide intake (Hogervorst et al., 2007). Acrylamide therefore
poses a potentially significant risk to human health.
Journal of Immunological Methods 341 (2009) 19–29
Abbreviations: ELISA, enzyme-linked immunosorbent assay; Ade, acryla-
mide; Hb, haemoglobin; AdeHb, acrylamide-adducted haemoglobin; HRP,
horseradish peroxidase.
⁎ Corresponding author. Tel.: +44 1517 948243.
E-mail address: andrew_jpreston@hotmail.com (A. Preston).
0022-1759/$ – see front matter © 2008 Elsevier B.V. All rights reserved.
doi:10.1016/j.jim.2008.10.003
Contents lists available at ScienceDirect
Journal of Immunological Methods
journal homepage: www.elsevier.com/locate/jim