Study On Biochip Technology And Fabric Imprint Protection Of Silk Archaeological Residues

China is the prior country to plant mulberry trees,raise silkworms and weave the silk textile in the world.The silk textile represents the highly developed scientific and technological civilization of the ancient China,which has made remarkable contributions to the development of the human civilization.Due to the lack of the sufficient archeological silk remains and the limitation of the detection methods,the research such as,the study on the application of silk fibers,the weaving origin of the silk textile,and the spread of silk civilization has always been in the focus of the academic study.Influenced by the adverse burial environmental factors,the silk fibres was easily degraded and even totally disappeared.But the fibroin residues were still survived and reserved in the soil.Therefore,the separation and identification of fibroin residues in the archaeological sites can provide the scientific evidence on the origin,the spread and the development of the ancient silk.similar as other kinds of the archaeological residues,the fibroin residues were also difficult to evaluate its preservation form and status with low abundance and complex contamination,which made it difficult in extraction,separation and purification of the fibroin residues.Athough,the biomass spectrum technology was often employed to identify the fibroin residues,the method requires much experimentaltrials for professionals,which greatly limits the application and promotion of the biomass spectrum technology.In this study,the form of the fibroin residues was firstly investigated,and the extraction technology of the fibroin residues was established and optimized,in order to obtain more information on the archaeological fibroin residuesBased on the structure and sequence characteristics of silk fiber and silk protein with the understanding of its degradation process,the characteristics of the residual peptide fragment was analyzed.Combining with the detection frequency of the peptide fragments,the antigen sequences of archaeological silk residues was established,and the monoclonal antibody was prepared and screened.On this basis,the detection macro array biochip technology of silk protein residues was developed,which was suitable for a rapid and accurate archaeological fibroin residue detection technology for the archaeologist.In addition,this paper studied the textile imprinting reinforcement technology,in order to provide technical support for the preservation of this form of cultural relics.In order to accurately distinguish the status of silk protein residues,this paper took the degraded samples and the nearly powdered samples unearthed from the archaeological sites as the research objects.The analytical technology such as electron scanning microscope(SEM),infrared spectroscopy(IR)and X-ray diffraction(XRD)were used to detect the samples and analyze the interaction behaviours among the silk fibre or its degradation products with the soil.The research illustrated that the silk fibre were degraded into the fibroin polypeptides in molecular level,and then became the organic matrix by the biological mineralization process and form the complex structure which contained the fibroin polypeptides and the mineral particles.Once this structure was formed,it prevented the further degradation of the fibroin polypeptides and ensured the long time preservation of the silk protein residues extraction which promote the identification of the fibroin residues..Meanwhile,this paper studied the pretreatment of the composite with fibroin polypeptide fragments and mineral particles before the extraction of the fibroin residues.It indicated that the samples treated with 10%acetic acid and EDTA-2Na were able to effectively release the fibroin polypeptides which bounded to the mineral particles.The number and coverage of the fibroin polypeptide fragments extracted from the samples were greatly increasedafter the samples were treated by the above method.In order to identify the fibroin residues qualitatively and rapidly,the monoclonal antibodies against the fibroin residues and the biochip detection of the fibroin residues were studied.In the preparation of monoclonal antibodies,the collecting and analyzing the results of previous studies about the identification of the archaeological fibroin residues was firstly surveyed.It was determined that the polypeptides occurred with highly frequency in the identification results was chosen as the hapten of the monoclonal antibody against the silk protein.Based on the analysis of the fine structure,the aggregation structure and the chain structure of the silk fibroin,it was found that the polypeptides occurred in high frequency were in the crystal structure where occupied in highest proportion with sufficient stability.The stability of that structure improves the ability of degradation resistance,which was the guarantee of the preservation of the silk residual.Meanwhile,the large amount of crystal structure increases the possibility of detection in the soil environment.Thus,the polypeptide fragments with-GX(A/S/Y/V)-as the basic sequence unit were determined to be the hapten.The hapten coupled with the carrier protein which was immunized by mice,and then 6 strains of monoclonal antibodies with high sensitivity were screened.On the basis of this study,the macro array biochip to identify the fibroin residue was developed Among the optimizing of the recognition criteria,the detection limits could be reached to 80 ppb of the fibroin polypeptides.It was shown that the biochip detection for the archaeological fibroin residues was accurate,rapid and convenient.Otherwise,the consolidation and seperation the soil which carried the silks residues and vestiges were also investigated.PVB was determined to consolidate the silk vestiges,ensured the effectiveness of the consolidation and ensured the surface stability of the silk vestiges,which did not affect the subsequent extraction and identification of the fibroin residues.The consolidated silk vestiges separatedwas benefit for the further conservation and display.In this paper,the study on the morphology of silk protein residues was the fundamental to explain the silk fiber and silk residues that can be retained in early tombs and sites,and was also the prerequisite for the extraction and identification of silk protein residues.The study on the extraction and pretreatment of silk protein residues can be used as a beneficial supplement to the whole extraction technology,and can be effectively improved the opportunities to obtain silk protein residues.The macro array biochip technology for silk protein residues detection will provide a convenient,fast and accurate qualitative detection technology for silk protein residues not only for front-line archaeologists,but also meet the increasing needs of archaeological identification.