Genetic immunization: a new monoclonal antibody for the detection of BCL-6 protein in paraffin sections.
Genetic immunization can be combined with hybridoma technology to generate high-affinity monoclonal antibodies (MAbs). A new anti-BCL-6 MAb (GI191E/A8) was produced by cloning full-length BCL-6 cDNA into a eukaryotic vector and delivering this into mouse epidermis using a helium gene gun. A comparative study was made of the specificity and the effects of formalin fixation on immunohistochemistry quality of GI191E/A8 and two other anti-BCL-6 MAbs. To evaluate its possible application to differential diagnosis of lymphomas, two tissue microarrays (89 diffuse large B-cell lymphomas and 24 B-cell chronic lymphocytic leukemia cases) were stained with GI191E/A8 and another anti-BCL-6 MAb produced by conventional means. Using GI191E/A8, the detection of BCL-6 protein was significantly increased, and its specificity was independent of formalin-fixation time. Using automatic quantified analysis, the correlation between the two anti-BCL-6 MAbs tested was identical in cases with overexpression or absence of BCL-6. In cases with intermediate BCL-6 protein expression, detection with GI191E/A8 was more sensitive. A significant association of higher BCL-6 expression and longer median overall survival times in diffuse large B-cell lymphomas was found. Using conventionally produced MAbs in the same patient group, the association was not significant.
García
José-Francisco
JF
Monoclonal Antibodies Unit, Biotechnology Program, Centro Nacional de Investigaciones Oncológicas (Spanish National Cancer Centre), C/Melchor Fernández Almagro 3, E-28029 Madrid, Spain.
García
Juan-Fernando
JF
Maestre
Lorena
L
Lucas
Elena
E
Sánchez-Verde
Lydia
L
Romero-Chala
Silvia
S
Piris
Miguel-Angel
MA
Roncador
Giovanna
G
D000818:Animals; D000911:Antibodies, Monoclonal; D045744:Cell Line, Tumor; D004268:DNA-Binding Proteins / Q000276:immunology / Q000378:metabolism*; D003937:Diagnosis, Differential; D005404:Fixatives; D005557:Formaldehyde; D006801:Humans; D007150:Immunohistochemistry; D015451:Leukemia, Lymphocytic, Chronic, B-Cell / Q000175:diagnosis / Q000378:metabolism; D008228:Lymphoma, Non-Hodgkin / Q000175:diagnosis / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D014066:Palatine Tonsil / Q000378:metabolism; D016612:Paraffin Embedding; D051560:Proto-Oncogene Proteins c-bcl-6; D016019:Survival Analysis; D046888:Tissue Array Analysis
D000911:Antibodies, Monoclonal; C084103:BCL6 protein, human; D004268:DNA-Binding Proteins; D005404:Fixatives; D051560:Proto-Oncogene Proteins c-bcl-6; D005557:Formaldehyde