The Establishment Of Rapid Monitoring Method On Microcystin-LR

With the increasing environmental pollution, the state of freshwater eutuophication is becoming more and more serious causing occurrence of toxic cyanobactreial blooms in various. The toxins produced by toxic cyanobactreial blooms polluting freshwater bodies, especially the source water and drinking water has become increasingly serious, which caused a great threat to human and animal health. Microcystin is a large class of cyanobacterial toxins, of which the most common and the most toxic cyanobacterial toxins is microcystin-LR (MC-LR). MC-LR is a low molecular weight cyclic peptide with multiple toxicity to organisms, which has been sent into the national drinking water health standards (GB5749-2006) with specified limits of1μg/L. MC-LR detection methods are many, but there are expensive, time-consuming and laborious, complex problem, therefore, to establish a fast, convenient, real-time, specific method of MC-LR detection has an important significance.This article obtained the pure product of MC-LR by Microcystis aeruginosa culture, and then collecting a sufficient number of algal cells for MC-LR extracted, through ultrasonic crushing, solvent extraction, and purified by high performance liquid chromatography (HPLC) separation, freeze-dried. Taking the MC-LR as target molecules, using phage display peptide library random screening to acquire MC-LR specific affinity ligand sequences. After four rounds screening, sequencing and sequence alignment, we ultimately obtained a MC-LR specific affinity ligand sequence. Then MC-LR specific affinity ligand peptide probe was synthesized by solid phase chemical synthesis method, and at the N-terminal with FITC labeled, and finally we researched the activity detection and applications of the prepared probe, validated and established a new method of biology.The experimental results show that the MC-LR extract obtained a purity of94%. After four round of the Ph.D.-12Phage Display Peptide Library Kit subsequently panning and sequence alignment, we got a MC-LR specific binding ligand sequence: H-F-F-K-W-H-T-R-T-N-D-Q,and then gained the MC-LR specific affinity ligand peptide P1with FITC labeled by artificial synthesis:FITC-Acp-H-F-F-K-W-H-T-R-T-N-D-Q-COOH, via fluorescent ELISA testing know that ligand peptide P1and MC-LR has a certain binding activity. Take peptide P1as a fluorescent probe to detect the actual water samples and compared with HPLC detection results, indicating that the method is strong sensitivity, higher specificity, and a fast, easy to operate, and low cost, can provide the new products and new methods of biology for the detection of water safety.