CD34+ stem cell counting using labeled immobilized anti-CD34 antibody onto magnetic nanoparticles and EasyCounter BC image cytometer.
Anal Biochem
; 610: 113929, 2020 12 01.
Article
en En
| MEDLINE
| ID: mdl-32866464
ABSTRACT
The ability of immobilized conjugate anti-CD34+ monoclonal antibody-dR110 and free conjugate anti-CD45+ monoclonal antibody-ATTO620 to precisely enumerate CD34+ stem cells and CD45+ cells in apheresis samples were evaluated. The conjugates anti-CD34+ antibody-dR110 and anti-CD45+- antibody-ATTO620 were prepared. Functionalized magnetic nanoparticles (MNPs) were synthesized. The anti-CD34+ antibody-dR110 conjugate was immobilized on the modified MNPs using a carbodiimide method. The stem cell count in thawed apheresis samples was determined using the free and the immobilized conjugate anti-CD34+ antibody-dR110 on MNPs and an image cell counter EasyCounter BC. A higher stem cell count and more accurate results were obtained with the immobilized conjugate, because a separation and concentration of the stem cells bound to antibody-dR110 on MNPs by external magnet were performed. Coefficients of variation of CD34+ cell count in apheresis samples, determined by EasyCounter BC, were ranged from 5.5 to 6.9% and those of CD45+ cell count from 3.8 to 4.7%. The viability of CD34+ cells was high from 98.5 to 99.6%. It was found that correlation coefficient between the flow cytometer and automatic cell counter, using free anti-CD34+ antibody-dR110 was 0.94, and when using immobilized anti-CD34+antibody-dR110 on MNPs, the correlation coefficient was 0.97.
Palabras clave
Texto completo:
1
Colección:
01-internacional
Base de datos:
MEDLINE
Asunto principal:
Antígenos CD34
/
Nanopartículas de Magnetita
/
Citometría de Flujo
/
Anticuerpos
Límite:
Humans
Idioma:
En
Revista:
Anal Biochem
Año:
2020
Tipo del documento:
Article
País de afiliación:
Bulgaria