A simple, computer-assisted assay to detect isotype-specific regulation of human immunoglobulin synthesis.
A simple, reliable, and computer-assisted assay has been developed to quantitate isotype-specific regulation of human immunoglobulin synthesis in vitro. The assay utilizes three separate human lymphoblast or myeloma cell lines, which secrete human immunoglobulins IgA, IgG, and IgE. Culture supernatants from 96-well tissue culture plates are then assayed for IgA, IgG, and IgE by a solid-phase enzyme-linked immunosorbent assay (ELISA) on a microtiter plate. Data collection and analysis is performed with the aid of computer programs designed for this assay. This assay has several advantages over other immunoglobulin regulation assays: no radioisotopes are used, thereby reducing cost and complexity; results are directly collected and quantified by computer analysis; the entire assay is completed in 3 days; reliability and reproducibility are increased by the use of established human cell lines; and co-culturing all three immunoglobulin-producing cell lines provides convenient internal controls for isotype specificity.
D002460:Cell Line; D002478:Cells, Cultured; D003513:Cycloheximide; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D006801:Humans; D007070:Immunoglobulin A / Q000032:analysis; D007073:Immunoglobulin E / Q000032:analysis; D007074:Immunoglobulin G / Q000032:analysis; D007136:Immunoglobulins / Q000096:biosynthesis* / Q000145:classification; D012636:Secretory Rate; D012984:Software; D013997:Time Factors
D007070:Immunoglobulin A; D007074:Immunoglobulin G; D007136:Immunoglobulins; D007073:Immunoglobulin E; D003513:Cycloheximide