Gruen L C, McInerney T L, Webster R G, Jackson D C
CSIRO Division of Biomolecular Engineering, Parkville, Victoria, Australia.
J Protein Chem. 1993 Jun;12(3):255-9. doi: 10.1007/BF01028188.
Sedimentation equilibrium centrifugation has been applied to determine the affinity and stoichiometry of the interaction between Fab fragments, derived from monoclonal antibodies NC10 and NC41, with influenza virus neuraminidase N9 isolated from either tern or whale. Although the two neuraminidase epitopes recognized by NC10 and NC41 Fab overlap, crystallographic studies have shown that the modes of binding of each Fab are different. The sedimentation equilibrium experiments described here reveal that the binding affinities are also different, with NC10 Fab binding more strongly to each neuraminidase. Furthermore, comparison of the affinity of binding of each antibody fragment reveals a stronger interaction with tern neuraminidase than with whale neuraminidase. Although the respective epitopes recognized by each antibody on the two antigens are similar, this technique shows that they do nevertheless possess sufficient differences to affect significantly the binding of antibody.
沉降平衡离心法已被用于确定源自单克隆抗体NC10和NC41的Fab片段与从燕鸥或鲸中分离出的流感病毒神经氨酸酶N9之间相互作用的亲和力和化学计量关系。尽管NC10和NC41 Fab识别的两个神经氨酸酶表位重叠，但晶体学研究表明每个Fab的结合模式不同。此处描述的沉降平衡实验表明，结合亲和力也不同，NC10 Fab与每种神经氨酸酶的结合更强。此外，比较每个抗体片段的结合亲和力发现，与燕鸥神经氨酸酶的相互作用比与鲸神经氨酸酶的更强。尽管两种抗原上每种抗体识别的各自表位相似，但该技术表明它们确实存在足够的差异，足以显著影响抗体的结合。
