Functional Compatibility Of Super-capacity Raman Barcodes For High-throughput Screening

Super-capacity optical barcodes may play an important role for high-throughput screening of biomolecular libraries.It is of great significant for the barcoding system to be compatible with the encoding synthesis,biomolecular synthesis and the functional screening processes.The encoding materials for the current optical barcoding systems mainly include organic dyes,quantum dots,upconversion nanoparticles and surfaceenhanced Raman scattering nanoparticles etc.The application of these barcoding system is limited by the chemical and optical stability of the encoding materials.Raman spectroscopy provides a solution for super-capacity spectral barcodes,because Raman signals have the advantages of being narrow-banded and stable.The small alkyne molecules in the Raman silent region and azo benzene molecules in the fingerprint region are excellent candidate for Raman encoding.In this thesis,in order to further expand the applicable band of and improve the chemical compatibility of the Raman barcoding system,we designed and synthesized libraries of Raman molecules including alkyne molecules with signal at frequencies of silent region and azo molecules at fingerprint region,and developed a super-capacity encoding system which can be compatible with highthroughput peptide screening.The main contents of this thesis are as follows:(1)We designed and synthesized a library of candidate Raman molecules chemical compatibility screening.The molecules were screened in the library synthesis condition and the surviving molecules were found.Using resin beads as a carrier,the Raman signal molecules were built up by these surviving molecules.The coding signals were tested and verified the compatible in the peptide library synthesis conditions.The construction of multiple sets of codes on a single plant was realized through the stepwise orthogonal deprotection strategy.The multiple sets of coding signals were tested and verified the compatible in the peptide library synthesis conditions.(2)The Raman barcoding system was used for high-throughput screening of combinatorial library of peptides.Use the split and pool strategy to build a combinatorial library of 10 ×10 ×10 OBOC peptides for screening a specific ligand that binds tumor cells,in which each peptide corresponds to a unique feature code.The library was incubated with U87-MG cells and the positive beads were decoded in situ in minutes.The high-throughput screening and decoding results indicated that all positive beads contain RGD sequence,and the LXW7 polypeptide containing GRGD is the most abundant.The Raman sequence encoding system established in this paper has good chemical compatibility,and improves the screening throughput of biologically active molecules in high-throughput screening.