The Study Of Lectin Modified Thermoresponsive Chromatographic Materials

In organisms,the abundance of glycoproteins is low.And the changes of glycoproteins are often relevant to the occurrence of many diseases,such as the cancer,rheumatoid arthritis,AIDS and so on.Lectin affinity chromatography is one of the most widely used methods in the separation and enrichment of glycoproteins.In recent years,the temperature sensitive chromatography has become a hot topic.The temperature sensitive chromatography can not only ensure the specific enrichment of the target molecules but also change the temperature to enrich the target molecules.The composition of mobile phase and pH value were not changed.In this paper,two kinds of controlled radical polymerization were used to synthesize the lectin modified temperature sensitive affinity chromatography materials.The glyproteins were enriched by the changing of the temperature.The graft density can be controled by the change of molar ratio of 2-bromide isobutyl bromide and acetyl chloride by ATRP.There were four kinds of graft density of the temperature sensitive materials by ATRP method,namely 10%,30%,50% and 70%.Then the NAS and NIPAAm were introduced.Their molar ratio was 1/10.Finally,the ConA was introduced to the surface of silica by the active intermediate NAS.The ovalbumin was used to analyze the performance of Silica@P(NIPAAm-co-ConA)with different grafting density.They were incubated at10 ℃ and 50 ℃ with different pH.The results showed that the Silica@P(NIPAAm-co-ConA)had the temperature sensitivity and enrichment performance.And the grafting density of 50% had a good performance at pH7.The release amount of the ovalbumin was 1.3 mg/g.The graft density of 50% temperature sensitive affinity chromatography was synthesized by RAFT.The monomer NAS and ConA were introduced on the surface of silica.The results of infrared spectrum,X-ray photoelectron spectroscopy and thermal gravimetric analysis showed that the synthesized products were in accordance with the experimental design.In HPLC,the stationary phase Silica@P(NIPAAm-co-ConA)had theperformance of capturing the hosphate buffer at 10 ℃.And the HRP can be released at50 ℃.The results showed that the lectin had been successfully introduced to the surface of silica.And the materials had a certain performance of temperature sensibility and entichment.