초록
2-브로모프로피오닐화된 수지를 이용해 insulin A (1-21) 사슬을 합성하였다. 시스테인의 결사슬은 각각 acetamidomethyl, benzyl, 그리고 benzhydryl기로 보호하였으며 그루타민과 아스파라긴은 p-nitrophenyl기로 활성화하여 합성에 이용하였다. 매 짝지음단계마다 DCC/HOBT coupling agent로 각 아미노산을 축합하였으며 반응의 완결여부는 닌히드린시험으로 측정하였다. 생성물은 NH$_3$/MeOH-Dioxane(v/v 1:1)로 수지로부터 분리하여 DEAE Sephadex A-25와 Sephadex LH-20으로 정제하였으며 최종생산물은 HPLC, electrophoresis로 확인한 결과 순수한 것(>99.9%)으로 나타났으며 총수득율은 6%이었다.
The total synthesis of insulin A chain (1-21) with properly protected sulfhdryl groups of three cysteins for the correct intra and inter disulfide bond formation has been accomplished on 2-bromopropionylated 2% DVB-styreneresin support employing manually operated rotary vessel. The sulfhydryl groups of cysteins were protected with acetamidomethyl, benzyl, and benzhydryl respectively. Glutamine and asparagine were attached to the peptide chain by active ester coupling, all other amino acids were coupled with DCC/HOBT. The synthesized peptide was purified by DEAE Sephadex A-25 and gel filtration Sephadex LH-20. The final product was found to be homogeneous by HPLC, electrophoresis, and amino acid analysis. The overall yield of the pure isolated peptide was 6%.
