Iron Oxide Based Serum Molecules And Circulating Tumor Cells Detection And Profiling

Mass spectrometry(MS)has been widely used for profiling and detection of diverse molecules considering its desirable analytical throughput,sensitivity,and capability of molecular identification and structural characterization.The characterization of circulating tumor cells(CTCs)plays a significant role in cancer prognosis,diagnosis and personalized treatment.In a case of CTCs,current full range of CTCs molecular characterization includes proteomic(e.g.immuno-fluorescence)and genetic strategies(such as fluorescence in situ hybridization and quantitative RT-PCR)to detect proteins and nucleic acids,respectively.Despite that,metabolism study of CTCs is largely hindered without a designer platform combining appropriate multiple analytical techniques.With the development of materials science,it plays a critical role in MS application and CTCs detection,but still faces a serious obstacle for further utilizing due to the surface chemistry and muti-functional application.Therefore,we prepared muti-functional ferric oxide particles for serum small molecules analysis and CTCs profiling with desirable efficiency and sensitivity.This work starts a facile designer technology for combining CTCs isolation and downstream metabolic analysis and provides new insights towards characteristics of rare cells in diverse dynamic biological process.This dissertation includes the following four parts:1.In chapter one,we offered a brief introduction to biological MS techniques.Then the methods of CTCs characterization and detection were reviewed.The development of material techniques were outlined and the emphasis is the ferric oxide particles application in MS and CTCs.In order to address the urgent challenges,we introduced our research points and novel ideas.2.In chapter two,we demonstrate a novel approach using pristine rough ferric oxide particles for LDI MS profiling and detection of serum small metabolites in bio-diagnostics.The highlights of this work includes: 1)highly sensitive LDI MS by the particles consuming 500 pL-500 nL of native serum without separation or enrichment in direct profiling of a variety of small metabolites;2)quantitation can be achieved for diverse small metabolites with enhanced detection-of-limit and dynamic ranges;3)a clinical demonstration in the measurement of serum glucose levels.3.In chapter three,several different types of functional magnetic particles were prepared for surface chemistry optimization in cell separation experiments.Further we evaluated the capture efficiency of the magnetic particles in PBS system and the whole blood system.For low cell concentrations,we optimized the particles to achieve desirable capture efficiency.Finally,the selected magnetic particles were characterized and loaded into a designed microfluidic system for CTCs isolation from blood samples.4.In chapter four,we demonstrated the combination of rare cell isolation and particles assisted LDI MS(PALDI MS)to detect the metabolism of captured CTCs for metabolic study based on functional ferric oxide magnetic particles.More importantly,these particles served as ideal matrix for LDI MS.Consequently,sensitive PALDI MS analysis can be achieved towards small metabolites from complex bio-mixture due to the unique selective LDI process.Further isotope assisted quantification was coupled to investigate small molecules metabolism of ~50 CTCs.This work starts a facile designer technology for downstream metabolic analysis of CTCs and provides new insights towards characteristics of rare cells in diverse dynamic biological process.