Development of radioimmunoassays on microplate: application for CA-GI and CA-Br tumor markers.
By taking advantage of a newly available microplate counter for radioactivity and the organic solvent-resistant, pigmented microplates, we have successfully established radioimmunoassays (RIA) for both CA-GI and CA-BR on microplate for routine clinical use. In the process of assay development, we found that both pigmented PicoPlate, made of acrylonitrile, and polystyrene Microlite 2 can be coated with antialpha fetoprotein (AFP) and antinerve growth factor (NGF) and used for setting up immunoassays for AFP and nerve growth factors. There were no problems following a test format of either competitive binding or sandwich design. Microlites 2 is recommended over PicoPlate because Microlites 2 is made of polystyrene, which is less expensive and separable into 8-well strips or even single wells. Single-well separation allows for the use of regular gamma counters in case Topcount is unavailable. We also found that the sensitivity of these tests was not significantly affected even though Topcount counts the weaker beta emissions. Similar dose-response curves could also be generated between original Biomira tube assays and assays using PicoPlate or Microlite 2 coated with protein antigens CA-Br and CA-GI. Excellent correlations were also obtained between the microplate assays and the Biomira tube assays for CA-GI and CA-Br using groups of serum specimens from cancer patients. We recommend the development of various RIAs on the microplate: it requires less reagents and less sample handling by the technologists and it can be essentially automated.
Wu
J T
JT
Department of Pathology, University of Utah Medical Center, Salt Lake City.
Erali
M
M
D000906:Antibodies; D015295:Antigens, Tumor-Associated, Carbohydrate / Q000032:analysis*; D001667:Binding, Competitive; D004797:Enzyme-Linked Immunosorbent Assay; D005069:Evaluation Studies as Topic; D006801:Humans; D009414:Nerve Growth Factors / Q000276:immunology; D011863:Radioimmunoassay / Q000295:instrumentation* / Q000379:methods / Q000706:statistics & numerical data; D012015:Reference Standards; D012680:Sensitivity and Specificity; D000509:alpha-Fetoproteins / Q000276:immunology
D000906:Antibodies; D015295:Antigens, Tumor-Associated, Carbohydrate; D009414:Nerve Growth Factors; D000509:alpha-Fetoproteins