INTEC   05402
INSTITUTO DE DESARROLLO TECNOLOGICO PARA LA INDUSTRIA QUIMICA
Unidad Ejecutora - UE
artículos
Título:
An unusual cysteine VL87 affects the antibody fragment conformations without interfering with the disulfide bond formation
Autor/es:
ATTALLAH, CAROLINA; AGUILAR, MARÍA FERNANDA; GARAY, A. SERGIO; HERRERA, FERNANDO E.; ETCHEVERRIGARAY, MARINA; OGGERO, MARCOS; RODRIGUES, DANIEL E.; ATTALLAH, CAROLINA; AGUILAR, MARÍA FERNANDA; GARAY, A. SERGIO; HERRERA, FERNANDO E.; ETCHEVERRIGARAY, MARINA; OGGERO, MARCOS; RODRIGUES, DANIEL E.
Revista:
MOLECULAR IMMUNOLOGY
Editorial:
PERGAMON-ELSEVIER SCIENCE LTD
Referencias:
Año: 2017 vol. 90 p. 143 - 149
ISSN:
0161-5890
Resumen:
The Cys residues are almost perfectly conserved in all antibodies. They contribute significantly to the antibody fragment stability. The relevance of two natural contiguous Cys residues of an anti-recombinant human-follicle stimulation hormone (rhFSH) in a format of single-chain variable fragment (scFv) was studied. This scFv contains 5 Cys residues: VH22 and VH92 in the variable heavy chain (VH) and VL23, VL87 and VL88 in the variable light chain (VL). The influence of two unusual contiguous Cys at positions VL87 and VL88 was studied by considering the wild type fragment and mutant variants: VL-C88S, VL-C87S, VL-C87Y. The analysis was carried out using antigen-binding ability measurement by indirect specific ELISA and a detailed molecular modeling that comprises homology methods, long molecular dynamics simulations and docking. We found that VL-C87 affected the antibody fragment stability without interfering with the disulfide bond formation. The effect of mutating the VL-C87 by a usual residue at this position like Tyr caused distant structural changes at the VH region that confers a higher mobility to the VH-CDR2 and VH-CDR3 loops improving the scFv binding to the antigen.