Characterization of the linker peptide of the single-chain Fv fragment of an antibody by NMR spectroscopy.
A comparison of the single-chain Fv fragment of the antibody McPC603 (scFv) with its corresponding unlinked Fv fragment has been carried out with 15N-edited NMR spectroscopy. The two Fv fragments adopt the same structure, indicating that the linker does not perturb the folding of the domains. This also directly demonstrates that folding in vivo (Fv fragment) and in vitro (scFv fragment) leads to the same structure. The main differences in the spectra of the uniformly 15N-labeled scFv and Fv fragments are due to signals of Gly and Ser from the linker peptide of the scFv fragment. The linker peptide has been mapped with NMR spectra of 15N-glycine- and 15N-glycine/15N-serine-labeled scFv fragments. The 15N T2 relaxation data indicate that the linker peptide is more flexible than the rest of the molecule.
Freund
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C
Department of Structural Research, Max-Planck-Institut für Biochemie, Martinsried, Germany.
Ross
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A
Guth
B
B
Plückthun
A
A
Holak
T A
TA
D005998:Glycine / Q000737:chemistry; D007128:Immunoglobulin Fragments / Q000737:chemistry*; D009682:Magnetic Resonance Spectroscopy; D011994:Recombinant Proteins / Q000737:chemistry
D007128:Immunoglobulin Fragments; D011994:Recombinant Proteins; C055966:immunoglobulin Fv; D005998:Glycine