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V raziskovalnem delu smo se ukvarjali s sintezo peptidov na trdnem nosilcu z uporabo Fmoc/tBu sintezne metode. Sintezo smo izvajali ročno z uporabo plastične siringe opremljene s frito, kjer smo vzpostavili reakcijske pogoje, ki smo jih nato prenesli na pretočni reaktor. Z uporabo ročne SPPS in pretočne SPPS smo sintetizirali tri različne peptide. Z ročno SPPS smo sintetizirali peptid H-LSPPRYP-OH ter peptid H- GLTFDEIQGLTYLQV-OH, s pretočno SPPS pa smo sintetizirali peptid H-LSPPRYP- OH ter peptid H-TIEQDYILILAVMHCSREPGYWKSRR-OH. S sintezo peptida H- LSPPRYP-OH smo preučevali razlike med uporabo ročne ter pretočne sinteze peptidov na trdnem nosilcu. Peptid H-TIEQDYILILAVMHCSREPGYWKSRR-OH smo sintetizirali kot enega od peptidnih fragmentov, z namenom nadaljne sinteze proteina dolgega 96 aminokisin z uporabo serin/treonin kemične ligacije. Vse sintetizirane peptide smo okarakterizirali s HRMS ter HPLC analizo. Ugotovili smo, da je pretočna SPPS veliko hitrejša od manualne SPPS, vednar dobimo manjše količine produktov, pri tem je tudi njihova čistost nižja. This master's thesis describes the synthesis of peptides on a solid support using the Fmoc/tBu synthesis strategy. The synthesis was carried out manually using a syringe equipped with a frit, where the reaction conditions were established, which were then transferred to the flow reactor. Using manual SPPS and flow SPPS, we synthesized 3 different peptides. The peptide H-LSPPRYP-OH and peptide H-GLTFDEIQGLTYLQV- OH were synthesised by using manual SPPS, and peptides H-LSPPRYP-OH and H- TIEQDYILILAVMHCSREPGYWKSRR-OH were synthesized by using flow SPPS. With the H-LSPPRYP-OH peptide synthesis, we studied the differences between the use of manual and flow synthesis of peptides on a solid support. The peptide H- TIEQDYILILAVMHCSREPGYWKSRR-OH was synthesized as one of the peptide fragments, with the aim of further synthesis of a 96 amino acid long protein using serine/threonine chemical ligation. All synthesized peptides were characterized by HRMS and HPLC analysis. We found that flow SPPS is much faster than manual SPPS, however resulted products in lower yield and purity. |
