Preparation Of Microarrays Of Cell Membrane Carbohydrate Identification And Its Initial Application Research

ObjectiveCarbohydrates,proteins and nucleic acids are three important molecules of vital movement. With the development of molecular biology and cytobiology, we began to know more biological functions of carbohydrates. Carbohydrates can participate vital movement by many forms, which have close relationships with many diseases, such as cancer and infection. For this reason, carbohydrates will become a topic of life science in the future.At present, the critical technology problems of glycobiology are structural analysis and synthesis method. Scientists are devoting themseives to breakthrough this. Function study of carbohydrate chains is focused on the study of regulation and recognition mechanism on cellular level.Lectins are one kind of proteins that can specificly bind to carbohydrates by non-covalent bonds. The greatest characteristic of lectins is to recognize the complicated structure of carbohydrates on cell membrane. The high specificity of lectins makes them to be the useful research tool in the study of biology and clinic. At present, there are many methods to detect carbohydrate chains by lectins. But most of them are not so good. They can't comprehensively detect carbohydrate chain in one time. Some foreign scholars make lectin microarrays to detect glucoproteins, but they can't capture cells. The process of purification of glucoproteins costs so much time and money and can't detect cells directly.Our experiment is to make a kind of lectin microarrays that can specificly capture the tissue cells according to the high-flux of microarrays. This kind of microarrays won't have to purify glucoproteins, so it costs much less and the manipulation are brief. We can detect the fluorescence intensity of lectin microarrays to analyze the structure of carbohydrate chains on cell membrane, which help us to get the global cognition of carbohydrate chain on cell membrane. As a result, the lectin microarrays are new technology platform to observe and detect the structure of carbohydrate chains on cell membrane.Methods1,Preparation of lectin microarraysTo study the modifying methods on film bases, to test the lectin fixation results, to study the ability of lectin microarrays to capture carbohydrate chains on cell membrane.2,Initial application of lectin microarrays.To study the ability and specificity of lectins microarrays to capture carbohydrate chain on cell membrane.3,Property analysis of lectin microarraysTo study the stability and sensitivity of lectin microarrays.Results1,Preparation of lectin microarraysWhen silicane solution pH is 7.2 , hydration time is 20 min, the aldehyde group-modified slide can capture cells much better than amino-group- modified slide. The optimal sample application concentration of lectin is 1.4mg/ml.2,Initial application of lectin microarraysThe lectin microarrays can specificly capture the carbohydrate chain on different tissue cells of mouse.3,Property detection of lectin microarraysThe stability of lectin microarrays are good. The optimal cell concentration of incubation is 1.8×10~6个/ml.ConclusionsIn this study , we make a kind of stable lectin microarrays that can accurately and specificly detect the carbohydrate chain on glycoproteins directly as well as the carbohydrate chain specificity on different tissue cell membrane. This kind of lectin microarrays are very stable and have high-flux, avoiding the complex structure analysis of carbohydrate .We can use the lectin microarrays to capture cells instead of abstraction glycoproteins, which will cost us less time and money. This kind of lectin microarrays is a new technology platform to observe and detect the structure of carbohydrate chains on cell membrane as well as a powerful technology reserve and research foundation to detect carbohydrate chain specificity on tumour cells by lectin microarrays.