Uniform labeling of a recombinant antibody Fv‐fragment with 15N and 13C for heteronuclear NMR spectroscopy
- 5 August 1991
- journal article
- Published by Wiley in FEBS Letters
- Vol. 287 (1-2), 185-188
- https://doi.org/10.1016/0014-5793(91)80047-7
Abstract
The expression of functional antibody fragments in Escherichia coli enables a detailed analysis by NMR spectroscopy. This is demonstrated with the uniform labeling of an Fv-fragment (25 kDa) comprising the antigen binding site of an anitbody against 2-phenyloxazolone with 15N and 13C. The antigen-complexed Fv-fragment was analysed for a potential assignment by heteronuclear multi-dimensional NMR spectroscopy. For almost all backbone amides 15N/1H crosspeaks and for 80% of them TOCSY crosspeaks were observed. In a 13C-edited-HCCH-2D experiment 17 out of 18 threonine spin-systems were identified. Thus detailed assignments are possible, but some amino acid specific labeling in addition to uniform labeling will be required for complete assignments of Fv-fragmentsKeywords
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