Preparation Of Monoclonal Antibodies Against Heavy Metal Zinc Ion And Development Blocking ELISA For Detection Of Zinc Ion

Heavy metal Zinc is an essential element in animals, Exists in a variety of animal foods.The heavy metal of human body content is the most of trace elements is Zinc, the content of Zinc up to 3 g as much involved in the synthesis and activation of the animal body 200 kinds of enzymes.It is the body's metabolism indispensable substances. Zinc in the body to promote the synthesis of protein by the division of cells in the body,growth and regeneration, thus affecting the growth and development of children and adolescents.Zinc deficiency can cause a range of physiological phenomenon of human disorders, many organs and tissues physiological dysfunction. Such as stunted growth, loss of appetite, poor mental development, while taste and vision will also be affected.The proportion of zinc and other elements in human body was important. When Zinc copper ratio was too large, prone to Hypertension and cardiopathy. It indicated herald advanced cancer when Zinc molydenum ratio was too large.People who intaked of Zinc excessively can cause zinc poisoning with symptoms such as vomition, diarrhea, damagement of the liver, kidneys, blood vessels and the heart, and even cause death. Thus the content of zinc in foods should be limited, so we need a fast and efficient method to detect the content of heavy metals in food and the environment zinc ions, in order to ensure our food security.In this paper, a new method of detection heavy metal zinc ion was developed with immunoassay, which compared to traditional physical and chemical analysis with time-saving, labor-saving, low cost, easy to carry, easy to operate and other advantages. Basis on analyzing the immunogenicity of Zinc, artificial antigen of Zn2+-ITCBE-BSA was synthesized successfully and identified with SDS-PAGE, ICP-AES. Identified heavy metal Zinc ions through the identification and preparation of monoclonal antibodies against Zn2+-EDTA was prepared with cell fusion and their immunological feature were identified with ic ELISA, IC50 value and cross-rectively. The protein concentration immunogen and coating antigen were 8.48 mg/m L and 9.23 mg /m L; Zn2+contents were 101.1?g /m L and 90.09?g /m L; indirect ELISA titer 1:1.28 × 104, blocking ELISA detection of Zn2+-EDTA half inhibitory concentration(IC50) was 26.85 ng /m L, and EDTA and other metal ions chelate no cross-reaction.Basis on the Zn2+-EDTA m Ab, the blocking ELISA for detecting Zn2+( Zn2+-kit) was established and its performances were measured with blocking ELISA for sensitivity, spiked measured for accuracy, between intra batch tests measured for accuracy, cross-reaction for specificity. The sensitivity of kit was 7.46 ng/m L, its accuracy and it had no cross-reactive with other metal. The coefficient of variation of the affix recovery test was less than 15% and meet the requirements of the standard,and it had no cross-reaction with other heavy metal chelates, and the shelf life of 4? was 6 months.