[Preparation and characterization of a directly labeled mouse anti-human CD14 monoclonal antibody ZCH-2F9-FITC].
To prepare fluorescein isothiocyanate (FITC) directly conjugated to monoclonal antibody (McAb) anti-human CD14, ZCH-7-2F9 (2F9-FITC).



After 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.



A 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).



2F9-FITC has been successfully prepared and it can be applied in diagnosis and differentiation of monoblastic leukemias.
Ning
Bo-tao
BT
Department of Hematology and Oncology, The Affiliated Children's HospitalìCollege of Medicine, Zhejiang University, Hangzhou 310003, China.
Tang
Yong-min
YM
Shen
Hong-qiang
HQ
Yang
Shi-long
SL
Chen
Ying-hu
YH
Song
Hua
H
Shi
Shu-wen
SW
Qian
Bai-qin
BQ
Xu
Wei-qun
WQ
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000276:immunology; D002478:Cells, Cultured; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate / Q000032:analysis / Q000138:chemical synthesis*; D005455:Fluorescent Antibody Technique; D006801:Humans; D007945:Leukemia, Lymphoid / Q000473:pathology; D015470:Leukemia, Myeloid, Acute / Q000473:pathology; D018950:Lipopolysaccharide Receptors / Q000276:immunology*; D051379:Mice; D008807:Mice, Inbred BALB C; D009000:Monocytes / Q000166:cytology
D000911:Antibodies, Monoclonal; D018950:Lipopolysaccharide Receptors; D016650:Fluorescein-5-isothiocyanate