Abstract
Immunology and Infection
使用 1-Cyano-4-二甲基亚胺四氟化物 （CDAP） 激活和结合可溶性聚糖
ERRATUM NOTICE
Important: There has been an erratum issued for this article. Read more …结合疫苗是疫苗学的显著进步。对于多糖结合疫苗的制备，多糖可以方便地功能化，并使用1-氰化物-4-二甲基丙胺四氟溴酸酯（CDAP）与疫苗载体蛋白连接，这是一种易于处理的氰化物试剂。CDAP 通过在 pH 7-9 中与碳水化合物羟基组发生反应来激活多糖。CDAP 的稳定性和反应性高度依赖 pH 值。由于CDAP的水解作用，反应的pH值在激活过程中也会降低，这使得良好的pH值控制成为可重复激活的关键。原始 CDAP 激活协议是在室温下以未缓冲的 pH 9 解决方案执行的。
由于在这种情况下（<3分钟）下快速反应，以及伴随快速CDAP水解的快速pH值下降，在短时间内快速调整和维持目标反应pH值是很困难。此处描述的改进协议在 0 °C 下执行，可减慢 CDAP 水解速度，并将激活时间从 3 分钟延长至 ±15 分钟。二甲基亚胺（DMAP）也被用作缓冲，在添加CDAP试剂之前，预先调整多糖溶液到目标激活pH值。反应时间越长，加上 CDAP 水解速度较慢，以及使用 DMAP 缓冲器，使得在整个激活过程的整个持续时间内保持激活 pH 值更加容易。改进的协议使激活过程不那么狂热，更易重复，更易于扩展。
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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