Analysis Of Snake Venom Mixture Based On Multivariate Analysis Technology

BackgroundSnake venom,as a natural and highly efficient biological protein gland secretion,is known as "liquid gold" for its high medicinal value.The composition of snake venom is very complicated.In fact,its toxicological mechanism is the result of the combined action of multiple active ingredients,but the pathological analysis of multiple active ingredients is still lacking.We use the high-throughput protein fluorescent dye method to discriminate and analyze seven snake venom proteins.While using machine learning to identify and analyze snake venom mixed samples,and exploring the venom of snake venom,we look forward to advancing artificial intelligence methods to analyze mixed samples,analysis of complex pharmacology and pathology.Objectives(1)A method was developed for the analysis of various complex samples of snake venom proteins and their mixtures with fluorescent dyes;(2)Developed a fluorescence array sensor based on multiplex analysis of differentially stained snake venom protein fluorescence staining.Methods(1)According to the six representative proteins detected(phospholipase A2,?-cobratoxin,cardiotoxin,hyaluronidase,thrombin,haemocoagulase),We seleced four fluorescent dyes(eosin Y,fluorescein isothiocyanate isomer I,sulforhodamine B,titan yellow)with universal properties,and mix a certed concentration of dyes and proteins.Then use fluorescence spectroscopy to measureand the fluorescence intensities of six proteins in four dyes,and recorded the changes in fluorescence intensity before and after protein addition.(2)Constructed a multivariate analysis fluorescence sensor array.Use the method of multivariate discriminant analysis to statistically analyze the data obtained from the sensor array,and use linear discriminant analysis(LDA)and hierarchical cluster analysis(HCA)to visualize the data.(3)Using the constructed multivariate analysis sensor array,analysed the different proteins at the same concentration,different proteins at different concentrations,different proteins at the same concentration gradient,multi-component protein mixtures,mixed proteins with different concentration gradients,and naturally occurring snake venom mixtures.Results(1)Six representative proteins(phospholipase A2,?-cobratoxin,cardiotoxin,hyaluronidase,thrombin,haemocoagulase)in four fluorescent dyes(eosin Y,fluorescein isothiocyanate isomer I,sulforhodamine B,titan yellow)all had unique fluorescent fingerprints,which can be identified and analyzed 100%;(2)By using LDA and HCA,LDA can clearly distinguish 6 species of snake venom proteins,and HCA can specifically observe the relationship between 6 species of snake venom proteins;(3)The constructed fluorescence sensing array could clearly distinguish different proteins at the same concentration,different proteins at different concentrations,different proteins at the same concentration gradient,multi-component protein mixtures and mixed proteins with different concentration gradients,the above experiments were repeated in 14 groups with an accuracy rate of 100%,and the detection limit was 10 ng/mL;(4)On the constructed fluorescence sensing array,seven kinds of snake venom mixtures naturally occurring in nature could be completely distinguished,and the relationship between the seven kinds of snake venom mixtures was clearly observed.Conclusions(1)Fluorescence sensor array based on fluorescent dye combined with snake venom protein to produce changes in fluorescence intensity shows good selectivity and specificity for various snake venom proteins;(2)Based on the principle that the dye combined with the snake venom protein caused the fluorescence intensity to change,the constructed fluorescent sensor could quickly and sensitively detect the type of snake venom protein,and showed good selectivity and specificity for the mixed snake venom protein.