Production and Application of APAAP Complex in Immunocytochemical and Immunohistochemical Staining (Pages: 23-30)

M. Naseri *, SM. Moazzeni * , AA. Pourfathollah ,


Introduction: The present study was undertaken to produce alkaline phosphatase anti- alkaline phosphatase (APAAP) complex and to use it in one of the most practical methods of determining the site of antigen in tissue or in cell, i.e. APAAP technique, and its comparison with similar foreign products. Materials and Methods: In this basic and applied study the secreted antibodies of the two hybridomas A1 G9 G3 and A1G8 F7 produced in tarbiat Modarres university were concentrated and purified and their affinities were determined. Then the complexes of monoclonal antibody against alkaline phosphatase and alkaline phosphatase enzyme (APAAP complex) were prepared in their optimal concentration and used in immunocytochemistry and immunohistochemistry staining. The results were compared with similar foreign sample (DAKO). Results: Both cell colonies had saved the potency of producing anti-alkaline phosphatase monoclonal antibody with high affinity. The produced APAAP complex was very effective and its sensitivity and specificity were comparable to the similar foreign kit (i.e. DAKO), Conclusion: The high affinity of the monoclonal antibodies of the two hybridomas and also the stability of the resulted complex from mixing monoclonal antibodies with the enzyme make it possible to use these two cell colonies to produce trading amounts of APPAP. The produced APAAP complex could be used in immunocytochemistry and immunohistochemistry staining.