Furlan M, Perret B A, Beck E A
Biochim Biophys Acta. 1979 Aug 28;579(2):325-33. doi: 10.1016/0005-2795(79)90060-6.
Human and bovine factor VIII were isolated from cryoprecipitate of fresh frozen plasma by gel filtration on Sepharose CL-2B. The elution diagrams and SDS-agarose electrophoretic analysis of eluted fractions show no significant differences in the size-distribution of factor VIII aggregates between the two species. Agarose gels were stained for carbohydrate by two methods: (1) the dansyl hydrazine reaction following oxidation with periodic acid and (2) staining with fluorescein-labeled concanavalin A. Results of both procedures indicate that in human factor VIII neither the size distribution nor its ristocetin cofactor activity are related to carbohydrate content. Bovine factor VIII contains slightly less sugar than the human preparation as judged from the relative dansyl hydrazine staining intensities. In contrast to human factor VIII, the binding affinity for concanavalin A of bovine factor VIII was gradually decreased with increasing aggregate size. This finding suggests an impaired accessibility of reactive sugar residues in large aggregates of bovine factor VIII.
通过在Sepharose CL - 2B上进行凝胶过滤，从新鲜冷冻血浆的冷沉淀中分离出人源和牛源凝血因子VIII。洗脱图以及洗脱级分的SDS - 琼脂糖电泳分析表明，两种物种的凝血因子VIII聚集体的大小分布没有显著差异。用两种方法对琼脂糖凝胶进行碳水化合物染色：（1）高碘酸氧化后进行丹磺酰肼反应，（2）用荧光素标记的伴刀豆球蛋白A染色。两种方法的结果均表明，人凝血因子VIII的大小分布及其瑞斯托霉素辅因子活性均与碳水化合物含量无关。从相对丹磺酰肼染色强度判断，牛凝血因子VIII含有的糖比人制品略少。与人凝血因子VIII不同，牛凝血因子VIII对伴刀豆球蛋白A的结合亲和力随着聚集体大小的增加而逐渐降低。这一发现表明，牛凝血因子VIII大聚集体中反应性糖残基的可及性受损。
