Abstract
Immunology and Infection
1-시아노-4-디메틸아미노피리딘 테트라플루오로보라산염(CDAP)을 이용한 용용다당류의 활성화 및 융합
ERRATUM NOTICE
Important: There has been an erratum issued for this article. Read more …공주 백신은 백신학에서 놀라운 발전입니다. 다당류 컨쥬게이트 백신의 제조를 위해 다당류는 1-시아노-4-디메틸아미노피리딘 테트라플루오로보레이트(CDAP)를 사용하여 백신 담체 단백질과 편리하게 기능화되고 연결될 수 있으며, 취급하기 쉬운 시안광과식 시약제입니다. CDAP는 pH 7-9에서 탄수화물 하이드록실 그룹과 반응하여 다당류를 활성화합니다. CDAP의 안정성과 반응성은 pH에 따라 매우 의존적입니다. 반응의 pH는 또한 CDAP의 가수분해로 인해 활성화 하는 동안 감소, 좋은 pH 제어 재현 가능한 활성화에 키를 만든다. 원래 CDAP 활성화 프로토콜은 버퍼링되지 않은 pH 9 솔루션의 실온에서 수행되었습니다.
이러한 조건하에서의 신속한 반응(<3분)과 빠른 cdAP 가수분해로부터의 빠른 pH 낙하로 인해 짧은 시간 내에 표적 반응 pH를 신속하게 조정하고 유지하는 것이 어려웠다. 여기에 설명된 개선된 프로토콜은 CDAP 가수분해를 늦추고 활성화 시간을 3분에서 ~15분으로 연장하는 0°C에서 수행됩니다. 디메틸아미노피리딘(DMAP)은 CDAP 시약을 첨가하기 전에 표적 활성화 pH에 대한 다당류 용액을 미리 조정하는 버퍼로 도용하였다. 느린 CDAP 가수분해 및 DMAP 버퍼의 사용과 결합된 긴 반응 시간을 통해 활성화 프로세스의 전체 기간 동안 활성화 pH를 보다 쉽게 유지할 수 있습니다. 향상된 프로토콜을 통해 활성화 프로세스는 열광적이고 재현가능하며 확장이 더 가능합니다.
Erratum
Erratum: Activation and Conjugation of Soluble Polysaccharides using 1-Cyano-4-Dimethylaminopyridine Tetrafluoroborate (CDAP)An erratum was issued for: Activation and Conjugation of Soluble Polysaccharides using 1-Cyano-4-Dimethylaminopyridine Tetrafluoroborate (CDAP). A figure was updated.
Figure 4 was updated from:
Figure 4: Representative results for CDAP activation of dextran. Typical standard curves for the (A) resorcinol/sulfuric acid and (B) TNBS assays. The assay results for dextran activated with 0.25 and 0.5 mg CDAP/mg dextran are shown. Glucose was used as the standard for the resorcinol assay. Dextran, in mg/mL, is divided by 100 kDa to give a molar concentration. The hydrazide concentration is determined using ADH as the standard and the results expressed as µM Hz. (C) Calculation of hydrazide: dextran ratios.The level of derivatization was calculated as hydrazides per 100 kDa of dextran to facilitate the comparison between polymers of different average molecular weights. The % weight ratio of g ADH/g dextran was calculated using a MW of 174 g/mole for ADH. Please click here to view a larger version of this figure.
to:
Figure 4: Representative results for CDAP activation of dextran. Typical standard curves for the (A) resorcinol/sulfuric acid and (B) TNBS assays. The assay results for dextran activated with 0.25 and 0.5 mg CDAP/mg dextran are shown. Glucose was used as the standard for the resorcinol assay. Dextran, in mg/mL, is divided by 100 kDa to give a molar concentration. The hydrazide concentration is determined using ADH as the standard and the results expressed as µM Hz. (C) Calculation of hydrazide: dextran ratios.The level of derivatization was calculated as hydrazides per 100 kDa of dextran to facilitate the comparison between polymers of different average molecular weights. The % weight ratio of g ADH/g dextran was calculated using a MW of 174 g/mole for ADH. Please click here to view a larger version of this figure.
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