Improved ELISA proliferation assay (EPA) for the detection of in vitro cell proliferation by a new Ki-67-antigen directed monoclonal antibody (Ki-S3).
We describe a simplified and improved proliferation assay based on a conventional ELISA system for the in vitro measurement of cellular proliferation (ELISA proliferation assay = EPA). The assay is based on a new monoclonal antibody (Ki-S3) to the proliferation-specific Ki-67-antigen and is carried out in 96-well microtiter plates using conventional immunoenzymatic methods. Ki-S3 is an immunoprecipitating monoclonal mouse IgG1 antibody, which recognizes a formalin-resistant epitope of the Ki-67 antigen. It can be used to measure proliferating cells in the cell cycle phases G1, S, G2 and M. In phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) the absorbance values obtained with the EPA show a statistically significant correlation to the number of Ki-S3 positive cells in simultaneously immunostained cytospin slides (r = 0.88). A direct comparison with [3H]thymidine labeling reveals the test to be an equally sensitive method for monitoring cellular proliferation (r = 0.91). This assay is an improved ELISA proliferation assay, which is easy to perform, does not require time-consuming pretreatments and avoids the hazards of radioactive isotopes.
Frahm
S O
SO
Department of Pathology and Hematopathology, University of Kiel, Germany.
Zott
B
B
Dworeck
C
C
Steinmann
J
J
Neppert
J
J
Parwaresch
R
R
D000818:Animals; D000911:Antibodies, Monoclonal / Q000276:immunology; D002455:Cell Division; D002478:Cells, Cultured; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D005069:Evaluation Studies as Topic; D005260:Female; D006801:Humans; D007553:Isotope Labeling; D019394:Ki-67 Antigen / Q000276:immunology*; D007963:Leukocytes, Mononuclear / Q000187:drug effects / Q000276:immunology; D008297:Male; D051379:Mice; D008934:Mitogens / Q000494:pharmacology; D010835:Phytohemagglutinins / Q000494:pharmacology; D012680:Sensitivity and Specificity; D013194:Staining and Labeling / Q000379:methods; D013936:Thymidine / Q000378:metabolism; D014316:Tritium
D000911:Antibodies, Monoclonal; D019394:Ki-67 Antigen; D008934:Mitogens; D010835:Phytohemagglutinins; D014316:Tritium; D013936:Thymidine