Studies On The Preparation For Anti-Hg²⁺ Monoclonal Antibody (McAb) And The Method Of ELISA Detection For Hg²⁺

Heavy metals are a kind of considerable contaminants that can lead to side effects even if they are present in extremely minute quantities.They are the most insidious pollutants because of their nonbiodegradable nature and ability to persist for long periods. Through the food chain,heavy metals may deposit into the humane body,which cause serious health problems,specifically cancer,and even transfer to the next generation on the genetic level.Mercury is one of the most toxic elements of heavy metals.Therefore,the quantitative analysis of tracing heavy metals is extremely important in environmental and agricultural food monitoring.The detection of mercury still depends on traditionary instruments.Compared with them,immunoassays have the characteristics of being quick, inexpensive,simple to perform,and reasonably portable,highly sensitive and selective,etc. So immunoassays have been focused in home and abroad in recent years.In our study,mercuric ions was coupled to Keyhole Limpet Hemocyanin(KLH)with glutathione to gain a complete antigen KLH-GLU-Hg²⁺.Several BALB/C mice with the helps of freund's complete and incomplete adjuvants were immunized with this immunoconjugate.After the immunization for five times,the antiserum of mouse was assessed by indirect ELISA,the OD values of Hg²⁺-loaded antigen(KLH-GLU-Hg²⁺)were much larger than those of none metal antigen(KLH-Hg²⁺).Then synthesizing and secreting anti-Hg²⁺McAb cell 2C3 was obtained after a series of steps of cell fusion,hybridoma screening and positive hybridoma subcloning,etc.At last,ascites fluids were produced in freund's incomplete adjuvant-primed BALB/C mice by intraperitoneal injection,its titer was above 1:51200.The hybridoma cell 2C3 was selected to be used for further study.By the study of the pH etc,five impact factors,a Ci-ELISA established.A curve regress was formed by the logarithm of inhibition ratio and the logarithm of the Hg²⁺concentration as two influencing facts,the two factors represent apparent linear correlation in the range of 1 ng/ml～1000 ng/ml The standard curve was y=0.2423x+0.0979,R² reaches 0.9835.This experiment will lay foundations for further studying of the fast test kits for detecting mercury,probing the way of ELISA method for correlation research.