A fully validated flow cytometry method to quantitatively analyze active rATG in human serum and its application in pharmacokinetic study for therapeutic drug monitoring.
J Pharm Biomed Anal
; 234: 115483, 2023 Sep 20.
Article
en En
| MEDLINE
| ID: mdl-37454500
ABSTRACT
Rabbit anti-thymocyte globulin (rATG) has been widely used to prevent graft-versus-host disease (GvHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The therapeutic window of rATG is narrow, and it may increase the risk of relapse, viral reactivation, delayed immune reconstitution and GvHD when overexposed or underexposed. Therefore, a reliable method for detecting the rATG concentration in human serum by flow cytometry was established and fully validated for therapeutic drug monitoring. In this method, Jurkat T cells were used to capture active rATG in human serum, and PE-labeled donkey anti-rabbit IgG was used as a secondary antibody. The method showed good specificity, selectivity and excellent linearity at concentration of 0.00300-20.0â¯AU/mL. The intra- and interday precision values were all within 20% at four concentration levels for the analyte. The stock solutions of rATG showed no significant degradation after storage at ambient temperature for 8â¯h and at -â¯80⯰C for 481 days. No significant degradation of rATG in serum was observed at ambient temperature for 6â¯h, during six freezethaw cycles and at -â¯80⯰C for at least 373 days. This method was fully validated and successfully applied to monitor active rATG concentration in serum of patients with haploid-identical hematopoietic stem cell transplantation.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Enfermedad Injerto contra Huésped
/
Suero Antilinfocítico
Límite:
Humans
Idioma:
En
Revista:
J Pharm Biomed Anal
Año:
2023
Tipo del documento:
Article
País de afiliación:
China