Preparation and characterization of a directly labeled mouse anti-human CD14 monoclonal antibody ZCH-2F9-FITC.
- Author:
Bo-tao NING
1
;
Yong-min TANG
;
Hong-qiang SHEN
;
Shi-long YANG
;
Ying-hu CHEN
;
Hua SONG
;
Shu-wen SHI
;
Bai-qin QIAN
;
Wei-qun XU
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Antibodies, Monoclonal; biosynthesis; immunology; Cells, Cultured; Flow Cytometry; Fluorescein-5-isothiocyanate; analysis; chemical synthesis; Fluorescent Antibody Technique; Humans; Leukemia, Lymphoid; pathology; Leukemia, Myeloid, Acute; pathology; Lipopolysaccharide Receptors; immunology; Mice; Mice, Inbred BALB C; Monocytes; cytology
- From: Journal of Zhejiang University. Medical sciences 2005;34(2):167-171
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo prepare fluorescein isothiocyanate (FITC) directly conjugated to monoclonal antibody (McAb) anti-human CD14, ZCH-7-2F9 (2F9-FITC).
METHODSAfter generation and purification, the purity and the murine immunoglobulin subtype of the antibody were evaluated with SDS-PAGE and multicolor flow cytometry (FCM). 2F9 McAb was directly labeled with FITC through modified Marsshall's method and the positive rate of the 2F9-FITC on different types of leukemic cells were compared with the standard CD14-FITC by FCM.
RESULTA large quantity of purified 2F9 McAb was prepared. The subtype of 2F9 was murine IgG1kappa. 2F9-FITC was successfully manufactured with A295/A280 ratio of 0.44. The positive cell percentages of 2F9-FITC and CD14-FITC on the monocytes were 84.50% and 90.08%, respectively, while those on lymphocytes were only 0.52% and 1.01%. There was no significant difference between the CD14 expressions with 2F9-FITC and CD14-FITC on each type of leukemia (n=23, t=0.922, P=0.367).
CONCLUSION2F9-FITC has been successfully prepared and it can be applied in diagnosis and differentiation of monoblastic leukemias.
