一种新的统计方法在优化抗血友病A因子免疫纯化中的应用。

Application of a new statistical approach to optimize the immunopurification of antihemophilia A factor.

作者信息

Bihoreau N, Layet S, Fontaine-Aupart M P, Paolantonacci P

机构信息

T.M. Innovation (Centre National de Transfusion Sanguine-Institut Mérieux, Les Ulis, France.

出版信息

J Chromatogr. 1993 Jan 29;612(1):49-56. doi: 10.1016/0378-4347(93)80366-c.

DOI:10.1016/0378-4347(93)80366-c

PMID:8454702

Abstract

Our aim was to optimize the immunopurification process of human factor VIII. This purification was performed using a mouse monoclonal anti-factor VIII light-chain antibody. Previous dissociation of the factor VIII-von Willebrand factor complex with CaCl2 led to a 50% increase of the factor VIII adsorption on the immunosorbent. The optimization of the elution step required the analysis of the effects of two parameters, pH and ionic strength, on four different responses: elution yield, concentration, specific activity and stability of factor VIII. For this purpose, a multifunctional method using Doehlert matrices for statistically designed experiments was applied. This methodology allowed us to obtain, with only seven experiments, a 60% increase of the elution yield and a two-fold increase of the specific activity of factor VIII.

摘要

我们的目标是优化人凝血因子VIII的免疫纯化过程。该纯化过程使用小鼠单克隆抗凝血因子VIII轻链抗体进行。先前用氯化钙解离凝血因子VIII - 血管性血友病因子复合物导致凝血因子VIII在免疫吸附剂上的吸附增加了50%。洗脱步骤的优化需要分析两个参数（pH和离子强度）对四种不同响应（凝血因子VIII的洗脱产率、浓度、比活性和稳定性）的影响。为此，应用了一种使用Doehlert矩阵进行统计设计实验的多功能方法。该方法仅通过七次实验，就使凝血因子VIII的洗脱产率提高了60%，比活性提高了两倍。