Toward the point-of-care testing of alkaline phosphatase in human serum.
Biosens Bioelectron
; 265: 116694, 2024 Dec 01.
Article
en En
| MEDLINE
| ID: mdl-39180828
ABSTRACT
This work addresses challenges of simple but reliable determination of enzyme alkaline phosphatase (ALP) in µL-sized serum samples. The single-tube assay and plate-based immunoassay were developed by using a commercial glucose test strip (GTS) to transduce analytical signal from the ALP + glucose-6-phosphate (G6P) reaction. The glucose released from G6P contributed to the flow of anodic charge Q20s through GTS. The interferences from serum matrix including those from native glucose were eliminated by devising a signal ΔQ20s = Q20s (total) - Q20s (matrix). The ΔQ20s was proportional to ALP activity within a linear range of 7.4-720 IU L-1 for assay (R2, 0.995) and 10-300 IU L-1 for immunoassay (R2, 0.983), which far exceeded a normal clinical range for ALP in adult human serum (30-150 IU L-1). Both methods required no auxiliary enzymes or labels, and were accurate (93-99 % signal recovery) and precise (RSD ≤10%). The assay was a simpler option because it required only one 20-min incubation step to determine ALP. The immunoassay involved three steps but it was still less laborious (by â¼70%) than a traditional enzyme-linked immunosorbent assay (ELISA) of ALP due to the substitution of ELISA detection antibody with GTS. The GTS-G6P-based methods bring ALP testing closer to the point-of-care locations, which is important considering the growing role of ALP as a prophylactic, therapeutic, and anti-inflammatory agent, and a marker of human diseases.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Técnicas Biosensibles
/
Fosfatasa Alcalina
/
Pruebas en el Punto de Atención
Límite:
Humans
Idioma:
En
Revista:
Biosens Bioelectron
Asunto de la revista:
BIOTECNOLOGIA
Año:
2024
Tipo del documento:
Article
País de afiliación:
Estados Unidos
Pais de publicación:
Reino Unido