The Application Of Molecular Imprinting Polymers: Using Which To Study In Biochemistry As Artificial Antibody And Synthesis Of Nano-chitosan Microsphere Recognizing Of Transmembrane Special Protein In Tumour

This dissertation is composed of two parts:the first part is concerned to the using protein imprinted polymer (PIP) as artificial antibody to study in biochemistry; the second part is concerned to the study of synthesizing nano chitosan by imprinting the specific peptides on transmembrane protein erythroblastosis B2 (ErbB-2) of tumor.In the firsr part, we used a new method for enrichment of the low-content cellular protein in high molecular weight by molecular imprinting. The template protein, bacterial cloned immunoglobulin binding protein (BiP), was selectively assembled with assistant recognition polymer chain (ARPC) from their library, which consisted of numerous limited length polymer chains with randomly distributed recognitions and immobilizing sites. The assemblies of protein and ARPC were adsorbed by polymeric beads and immobilized by cross-linking polymerization. After the template was removed, the synthesized imprinted polymer was used to adsorb authentic BiP from endoplasmic re-ticulum (ER) extract, whose proportional content was then enriched 45 times. It is the first time that the low-content cellular natural protein with relatively high molecular weight (78 kDa) was enriched by molecular imprinted polymer. In the previous study of our group, we found that FK506-binding protein (FKBP23) could bind to immunoglobulin-binding protein (BiP) by biological method co-immunoprecipitation. Here this result was confirmed again by chemical method using PIP instead of co-immunoprecipitation. In addition, these two methods both confirmed that the binding between BiP and FKBP23 was regulated by [Ca2+] and that the binding could not be detected at a high [Ca2+]. Therefore, this new type of PIP provided a feasible alternative to biological antibodies in biochemical research.In the second part, specific peptides from ERBB-2, many different tumors were motivated by which, were bond onto glass sheet by using OTCS (Octadecyl trichlorosilane). Then, nano-chitosan was synthesized by using different methods without pepetide. The TEM of the particles showed that the polychitosan-acrylic acid microspheres was 50-150 nanometer, which meeted our demand. Finally, nano-chitosan microspheres in the presence of peptides on glass sheet and ARPC were synthesized. These will be a foundation for the further research on targeted therapy by nano-chitosan.