Preparation and Properties of Monoclonal Antibody against Heavy Metal Cd2+

doi:10.7506/spkx1002-6630-201321050

Abstract

Abstract:

Bifunctional chelator isotrhiocyanobenzyl-EDTA (iEDTA) was coupled with Cd2+ and conjugate with
ovalbumin (OVA) and bovine serum albumin (BSA) to obtain Cd-iEDTA-OVA as an immune-antigen and Cd-iEDTABSA
as a screening antigen, respectively. Cd-iEDTA-OVA was injected to BALB/c mice to produce mAb against Cd2+, and
Cd-iEDTA-BSA was utilized to screen the monoclonal antibodies (mAb) by enzyme-linked immunosorbent assay (ELISA).
Results showed that specific hybridoma cell lines 7F4 and 7E8 were obtained after immunizing mice with Cd-iEDTA-OVA containing
174.6230 μg/L Cd2+ and 1.7892 mg/mL OVA, and screening antibody with Cd-iEDTA-BSA containing 48.1881 μg/L Cd2+ and 1.8065 mg/mL
BSA. Five high purity mAbs were obtained, such as 7E8E5, 7E8G9, 7F4B8, 7F4D6 and 7F4H8, whose subtypes belong to
IgG1. Among them, the titer of 7E8E5 was the highest, which was 1:512000; the affinity constant (Ka) of 7E8E5 was
4.55 × 108 L/mol. As a result an indirect competitive ELISA method for Cd2+ was established, and the IC50 and detection
limit were 1150 ng/mL and 260 ng/mL (R2 = 0.9916), respectively. However, it showed strong cross reactivity with Hg2+,
weak cross reactivity with Zn2+, Cu2+ and Ca2+, and almost no cross reactivity with Mg2+, Ni2+, Al2+, Pb2+ or Ba2+.

Key words: cadium, antigen, hybridoma cell, monoclonal antibody (mAb), enzyme linked immunosorbent assay (ELISA)

CLC Number:

Q785

YI Cui-ping1，SU Fang1，CHEN Yong-fa1，PENG Xin-kai2，ZHU Xiang-rong3. Preparation and Properties of Monoclonal Antibody against Heavy Metal Cd2+[J]. FOOD SCIENCE, 2013, 34(21): 248-253.

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