RESUMO
The aim of this study is to develop creatine kinase isoenzyme MB (CK-MB) specific monoclonal antibodies (mAb), and characterize the monoclonal antibody and further development of quantitative detection assay for CK-MB. The BALB/c mice were immunized with purchased CK-MB antigen, then monoclonal antibodies were prepared according to conventional hybridoma technique and screened by indirect and capture ELISA method. To identify the epitopes and evaluate the classification, purchased creatine kinase isoenzyme MB (CK-MM/BB/MB) antigen was used to identify the epitopes, with immunoblotting and synthetic CK-MM and CK-BB in different linear epitope. A double antibody sandwich ELISA was applied to screen the mAb pairs for CK-MB detection, and the quantitative detection assay for CK-MB was developed. We used 74 cases of clinical specimens for comparison of our assay with Roche's CK-MB assay. We successfully developed 22 strains of hybridoms against CK-MB, these mAbs can be divided into linear, partial conformational CK-MB, CK-MM or CK-BB cross monoclonal antibody and CK-MB specific reaction with partial conformational monoclonal antibody, and CK-MB quantitative detection assay was developed by using partial conformational monoclonal antibody. The correlation coefficient factor r of our reagent and Roche's was 0.930 9. This study established a screening method for CK-MB partial conformational specific monoclonal antibody, and these monoclonal antibodies were analyzed and an established quantitative detection assay was developed. The new assay had a high concordance with Roche's.
RESUMO
<p><b>OBJECTIVE</b>To study the clinicopathologic and immunohistochemical features of diffuse large B-cell lymphoma (DLBCL) and the significance of immunohistochemistry in diagnosis and differential diagnosis of DLBCL.</p><p><b>METHODS</b>60 cases of DLBCL were studied and immunohistochemical staining for LCA, L26, BLA-36, CD30, bcl-6 were carried out with the EnVision 2 step method.</p><p><b>RESULTS</b>The age range of 76.7% (46/60) patients was 40 - 70 years. The location of the lesion includes nodal and extranodal sites. 90.0% (54/60) were in clinical stages of II (24/54), III (21/54), IV (9/54). Histopathologic morphology presented as centroblastic (88.3%, 53/60), immunoblastic (3.3%, 2/60), anaplastic large B cell type (3.3%, 2/60) and T cell rich B cell type (5.0%, 3/60). Immunostaining showed 100% (60/60) DLBCL were positive for LCA, L26, BLA36, 3.3% (2/60) DLBCL positive for CD30, 95% (57/60) expressed bcl-6 protein.</p><p><b>CONCLUSIONS</b>DLBCL is an aggressive lymphoma which shows cytologic variability from case to case. The evaluation of pathologic features and immunohistochemistry in DLBCL are useful and practical for diagnostic purposes, but cannot delineate distinctive morphologic subtypes.</p>