RT Journal Article
T1 Oriented immobilization of antibodies through different surface regions containing amino groups: Selective immobilization through the bottom of the Fc region
A1 Gao, Shipeng
A1 Rojas-Vega, Francisco
A1 Rocha Martín, Javier
A1 Guisán, José
AB Amino groups on the antibody surface (amino terminus and Lys) are very interesting conjugation targets due to their substantial quantities and selectivity toward various reactive groups. Oriented immobilization of antibodies via amino moieties on the Fc region instead of the antigen-binding fragment (Fab) is highly appreciated to conserve antigen-binding capacity. In this paper, targeting amino moieties on distinct regions, three antibody immobilization strategies were compared with the recognition ability of corresponding adsorbents. Our results demonstrate that oriented immobilization of antibodies onto heterofunctional chelate-epoxy support selectively involving Lys residues placed at the bottom of the Fc region, thus preserved the highest antigen recognition capacity (over 75% functionality). For homofunctional aldehyde support, immobilization at pH 10 demonstrates 50% remaining functionality due to the random orientation of tethered antibodies; while only 10% functionality remained when N-terminus were specifically conjugated at pH 8.5. With the rationalization of moieties density onto heterofunctional support, 2-fold recognition capacity was exhibited over randomly immobilization for antigens with higher size (β-galactosidase, 425 kDa vs. horseradish peroxidase, 40 kDa). Meanwhile, at least 97% of antigens with a varied concentration in diluted human serum were efficiently captured by the optimized chelate-epoxy support. Therefore, our antibody immobilization protocol proved the potential to be utilized as a promising candidate to capture voluminous antigens (large proteins and cells) in real samples.
PB Elsevier
SN 0141-8130
YR 2021
FD 2021
LK https://hdl.handle.net/20.500.14352/94115
UL https://hdl.handle.net/20.500.14352/94115
LA eng
NO Gao, Shipeng, et al. «Oriented Immobilization of Antibodies through Different Surface Regions Containing Amino Groups: Selective Immobilization through the Bottom of the Fc Region». International Journal of Biological Macromolecules, vol. 177, abril de 2021, pp. 19-28. https://doi.org/10.1016/j.ijbiomac.2021.02.103.
NO Este trabajo fue apoyado por el gobierno regional de Madrid ( Doctorados Industriales en la Comunidad de Madrid , IND2018/BIO-9480 ). Francisco Rojas-Vega desea agradecer al gobierno regional de Madrid por el apoyo financiero ( IND2018/BIO-9480 ). Shipeng Gao también agradece el apoyo financiero del Consejo de Becas de China ( 201808440415 ).
NO Comunidad de Madrid
NO China Scholarship Council
DS Docta Complutense
RD 6 abr 2025