Conjugation site modulates the in vivo stability and therapeutic activity of antibody-drug conjugates.
Nat Biotechnol
; 30(2): 184-9, 2012 Jan 22.
Article
en En
| MEDLINE
| ID: mdl-22267010
ABSTRACT
The reactive thiol in cysteine is used for coupling maleimide linkers in the generation of antibody conjugates. To assess the impact of the conjugation site, we engineered cysteines into a therapeutic HER2/neu antibody at three sites differing in solvent accessibility and local charge. The highly solvent-accessible site rapidly lost conjugated thiol-reactive linkers in plasma owing to maleimide exchange with reactive thiols in albumin, free cysteine or glutathione. In contrast, a partially accessible site with a positively charged environment promoted hydrolysis of the succinimide ring in the linker, thereby preventing this exchange reaction. The site with partial solvent-accessibility and neutral charge displayed both properties. In a mouse mammary tumor model, the stability and therapeutic activity of the antibody conjugate were affected positively by succinimide ring hydrolysis and negatively by maleimide exchange with thiol-reactive constituents in plasma. Thus, the chemical and structural dynamics of the conjugation site can influence antibody conjugate performance by modulating the stability of the antibody-linker interface.
Texto completo:
1
Bases de datos:
MEDLINE
Asunto principal:
Sitios de Unión de Anticuerpos
/
Inmunoglobulina G
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Ingeniería de Proteínas
/
Inmunoconjugados
/
Anticuerpos
Límite:
Animals
/
Humans
Idioma:
En
Revista:
Nat Biotechnol
Asunto de la revista:
BIOTECNOLOGIA
Año:
2012
Tipo del documento:
Article
País de afiliación:
Estados Unidos