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Solid-phase Synthesis and Purification for Fusukang

Author: ZhaoJiong
Tutor: GanYiRu
School: Tianjin University
Course: Biochemical Engineering
Keywords: Solid-phase synthesis Fragment condensation Fu Su Kang Ion exchange chromatography Reversed-phase high performance liquid chromatography
CLC: TQ463
Type: Master's thesis
Year: 2006
Downloads: 205
Quote: 3
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Abstract


This paper studies the 36 peptide of Fu Su Kang Fmoc solid- phase chemical synthesis . Fu Su Kang Fmoc solid phase step - by - step chemical synthesis and Fmoc solid phase chemical synthesis of fragment condensation . Fmoc solid phase of step - by - step chemical synthesis process conditions : DMF as the reaction solvent ; HBTU / HOBt / DIEA for condensing agent ; Wang resin is a solid phase carrier ; TFA/p- methylphenol / EDT / water ( 97/0.25/0.25 / 2.5, V / V) for the cutting conditions , the protected amino acid 2-fold excess of the condensation reaction. Solid phase fragment condensation the chemical synthesis Rotary prime Kang , synthesis method for the synthesis of the fully protected peptide fragment : 2 - chlorotrityl resin and Wang resin is a solid phase carrier ; HBTU / HOBt / DIEA as a condensing agent ; DMF The reaction solvent , HOAc / TFE / DCM (1:2:7, volume ratio) as the cutting conditions . Fragment condensation polypeptide fragment segmentation strategy the fragment condensation solvent , fragment condensation agent be reduced to pieces of legitimate synthetic Fu Su Kang quality , established the optimization of fragment condensation process conditions : Fu Su Kang residual group 1 to 10,11 to 20,21 to 25,26 to 36 divided into four segments; to Wang resin as the solid phase carrier ; HATU / of HOAt / DIEA as a condensing agent ( a 4-fold excess ); DMF / DCM (1:1 after completion of the condensation and deprotection ) is a reaction solvent ; fully protected peptide fragment per step added to 1.5 -fold excess ; washing solvent is washed with DCM for 5 times, the condensation reaction of the solid phase sheet segment . Synthesis under the conditions Fu Su Kang , crude yield of 67.31% , 38.75% purity by quantitative analysis of the standard curve . Fu Su Kang crude separation and purification , to establish a purification process route : preliminary purification using ion exchange chromatography , the final purification is prepared in the reversed-phase high performance liquid chromatography . By ion exchange chromatography , the purity of the product to 78% , the yield was 55% ; final purification of the results of a purity of 95% . The total yield of the two-step purification was 35 %.

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