Comparison of the amide proton exchange behavior of the rapidly formed folding intermediate and the native state of an antibody scFv fragment

FEBS Letters

Holak, Tad A²  Gehrig, Peter¹  Freund, Cristian¹  Plückthun, Andreas¹ 

[1] Biochemisches Institut, Universität Zürich, Winterthurerstr. 190, CH-8057 Zürich, Switzerland;Max-Planck-Institut für Biochemie, D-82152 Martinsried, Germany

关键词: scFv fragment;    H/D exchange experiment;    Mass spectrometry;    Nuclear magnetic resonance;   

DOI  :  10.1016/S0014-5793(97)00306-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

We have investigated the stability of backbone amide protons of the intermediate and the native state of the scFv fragment of an antibody. Stopped flow experiments analyzed by MS and NMR detected the formation of an exchange protected intermediate within the deadtime of the stopped flow apparatus (17 ms). H/D exchange rates of the native protein identified a number of very stable backbone amide protons in the V_L and the V_H domains. In the V_L domain, this slowly exchanging core of the scFv fragment is similar to the folding core of the intermediate, while the V_H domain possesses a great number of very stable amide protons which are not stabilized to a significant degree in the folding intermediate of the scFv fragment.

【 授权许可】

Unknown