Stable Isotope Iodine-labeled Antibodies-ICP-MS For Immunoassay

Immunoassay has been developed continually following the life science progressing.Recently,the Inductively Coupled Plasma Mass Spectrometry has gotten recognition because of it's outstangd advantage,such as,high sensitive,low detection limit,multi-component analysis ability and so on,so the immunoassay with high flux has a good foreground.In the study,the technique of stable isotope iodine labelling protein and ICP-MS linked immunoassay has been established,which was used in the dectetion of speciality of protein and bacteria.This dissertation consists of four chapters:1.Review;2.Iodine of stable isotope labelling protein;3.Non-radioactive Iodine-labeled Antibodies-Inductively Coupled Plasma Mass Spectrometry for Immunoassay;4.Vapour generation inductively coupled mass spectrometry in determination of iodineChapter one of this thesis was an overview of the development about immunoreaction and the field in which it was used;the labelling skills and development have been expatiated briefly,in addition,the standardization of labelling-Immunoassay and the developmental trend were discussed;especially,poping out the method of immunoassay linked with ICP-MS.In chapter two,the method about iodine of stable isotope labelling protein was established.The methods and the developments of different iodine-labelling ware introduced,and compared each other.During this study,the protein was tagged with non-radioactive ¹²⁷I using N-Bromosuccinimide(NBS)to produce the I-Compound:NBS is a kind of oxidant which is mild and the reaction happened in solution,so the labelling method was a perfect one which was convenient operation,high labelling tag-yield and the I-Compound being stable.In chapter three,the immunoassay about Iodine labelling protein-Inductively Coupled Plasma Mass Spectrometry was dissertated.Firstly,the goat anti-Escherichia Coli(GAE)and goat anti rabbit(GAR)were selected as the antigen and antibody respectively in the establishment of this immunoassay,and the iodine was detection by ICP-MS after reaction.In addition,the ornament of the polystyrene microtitre plate has been investigated;the method about disposal of sample was established,by which not only avoiding the volatilization of iodine and abating the effect of memory by ICP-MS.In the chapter,we also did some experiment about the detection of bacteria in the method.In chapter four,An online vapour generation ICP-MS method for the determination of iodine was established.In the study,depending on the deoxidization of the I^- and the means of oxidization,sensitivity was raised due to improving transfers efficiency to ICP-MS by using the technique which the I^- was oxidized to I₂ that was volatile before being detected.In addition,the different modality of iodine have been detected by the method.The main innovation of this thesis is:1.The non-radioactive Iodine(127)-labelled protein was established in order to combine ICP-MS with immunoassay.In the experiment,we adopted the method of NBS to achieve the protein labelling with iodine,the choice of stable iodine avoids radioactive iodine-labelling which was difficult for experiment because of the radioactivity.2.A new immunoassay was established by the ¹²⁷I labelling protein linked with ICP-MS,and the immunoassay was used in the detection of bacteria.3.An online vapour generation ICP-MS method for the determination of iodine was achieved,which the sensitivity can be raised due to improving transfers efficiency to ICP-MS by using Chemical Vapour Generation.