[Preparation and application of monoclonal antibodies against Heavy Metal Chromium]
Lu Yan1, Yong Tang, Jun-Jian Xiang
1Antibody Engineering Center, Jinan University, Guangzhou 510632, China. sylx66@yahoo.com.cn
Aim:
To produce the monoclonal antibodies (mAb) against Heavy Metal Chromium, and develop a competitive inhibition enzyme-linked immunosorbent assay (ciELISA) for the detection of Chromium.
Methods:
Chromium ions was chelated with bifunctional chelating agent Isothio-cyanobenzyl- EDTA firstly and then conjugated with bovin serum alburmin(BSA) and ovalbmin(OVA) respectively to complete antigens . Four female BALB/c mice were immunized with Cr-iEDTA-BSA, the Hybridoma lines secreting monoclonal antibodies against Chromium ions were established by the hybridoma technology. The titer and the specificity of the antibodies were characterized in the way of indirect enzym-elinked immunosorbent assay (iELISA) and ciELISA.
Results:
After immunization and cell fusion, three hybridomaes which can stably secrete mAbs against chromium were obtained. The antibody titer was up to 1×10(5);, only had 9.62% cross-reactivity to Fe(3+); and little to other ions. Linear detection of ciELISA for Chromium covered a range from 0.783 μg/L-50 μg/L with the limit at the level of 1 μg/L. An excellent correlation of results obtained by ciELISA and ICP-AES when the concentration more than 1 μg/L.
Conclusion:
The monoclonal antibody against Chromium with high sensitivity and specificity has been generated and a competitive inhibition enzyme-linked immunosorbent assay for the detection of Chromium ions has been established successfully, which was met the the national standard of water quality nonitoring.
