Preparation and characterization of a directly labeled mouse anti-human CD14 monoclonal antibody ZCH-2F9-FITC / 浙江大学学报·医学版
Journal of Zhejiang University. Medical sciences
; (6): 167-171, 2005.
Article
em Zh
| WPRIM
| ID: wpr-353225
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WPRO
ABSTRACT
<p><b>OBJECTIVE</b>To prepare fluorescein isothiocyanate (FITC) directly conjugated to monoclonal antibody (McAb) anti-human CD14, ZCH-7-2F9 (2F9-FITC).</p><p><b>METHODS</b>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.</p><p><b>RESULT</b>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).</p><p><b>CONCLUSION</b>2F9-FITC has been successfully prepared and it can be applied in diagnosis and differentiation of monoblastic leukemias.</p>
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Índice:
WPRIM
Assunto principal:
Patologia
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Monócitos
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Leucemia Mieloide Aguda
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Leucemia Linfoide
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Células Cultivadas
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Imunofluorescência
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Fluoresceína-5-Isotiocianato
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Receptores de Lipopolissacarídeos
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Biologia Celular
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Alergia e Imunologia
Limite:
Animals
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Humans
Idioma:
Zh
Revista:
Journal of Zhejiang University. Medical sciences
Ano de publicação:
2005
Tipo de documento:
Article