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Original ArticleOpen Access
Production of Monoclonal Antibody to CD4 Antigen and Development of Reagent for CD4+ Lymphocyte Enumeration
Kasinrerk W 
,
Tokrasinwit N ,
Naveewongpanit P
,
Tokrasinwit N ,
Naveewongpanit P A hybridoma secreting monoclonal antibody (mAb) specific to CD4 protein was generated.
This monoclonal antibody, named MT4, was proved to be specific to CD4 protein as it reacted with
CD4-DNA transfected COS cells, CD4+ cell line:; and CD4+ lymphocytes. Furthermore, MT4
mAb inhibited the binding of standard CD4 monoclonal antibodies to CD4 proteins on CD4+ cells.
To develop a home made reagent for CD4+ lymphocyte determination by flow cytometry, fluorescein
isothiocyanate (FITC) was conjugated to MT4 mAb. To evaluate the developed reagent,
30 HIV infected and 30 healthy individuals were determined for CD4+ lymphocytes by using
both a commercial Simultestâ„¢ reagent kit and home made FITC labeled MT4 mAb simultaneously.
The study has shown that both percentages and absolute CD4+ lymphocyte counts obtained
from both reagents were equivalent. The correlation coefficient for regression analysis was 0.995
and 0.996 for percentages and absolute CD4+ lymphocyte counts, respectively. The results suggest
that home made FITC labeled MT4 reagent is an acceptable alternative reagent for monitoring
CD4+ lymphocytes in blood samples by flow cytometry.
This monoclonal antibody, named MT4, was proved to be specific to CD4 protein as it reacted with
CD4-DNA transfected COS cells, CD4+ cell line:; and CD4+ lymphocytes. Furthermore, MT4
mAb inhibited the binding of standard CD4 monoclonal antibodies to CD4 proteins on CD4+ cells.
To develop a home made reagent for CD4+ lymphocyte determination by flow cytometry, fluorescein
isothiocyanate (FITC) was conjugated to MT4 mAb. To evaluate the developed reagent,
30 HIV infected and 30 healthy individuals were determined for CD4+ lymphocytes by using
both a commercial Simultestâ„¢ reagent kit and home made FITC labeled MT4 mAb simultaneously.
The study has shown that both percentages and absolute CD4+ lymphocyte counts obtained
from both reagents were equivalent. The correlation coefficient for regression analysis was 0.995
and 0.996 for percentages and absolute CD4+ lymphocyte counts, respectively. The results suggest
that home made FITC labeled MT4 reagent is an acceptable alternative reagent for monitoring
CD4+ lymphocytes in blood samples by flow cytometry.
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