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A screening method to detect clonal secretion of DNP-specific antibody
Journal of Cellular Physiology
|October 1, 1975
Summary
Researchers identified spontaneous variants of IgA-producing mouse myeloma cells (S194-2) that lost immunoglobulin secretion. This study focused on IgA variants lacking altered DNP binding sites or secreted subunits.
Area of Science:
- Immunology
- Cell Biology
- Biochemistry
Background:
- The S194-2 mouse myeloma cell line secretes IgA immunoglobulin with DNP-binding activity.
- Understanding immunoglobulin secretion variants is crucial for myeloma research.
Purpose of the Study:
- To isolate and characterize spontaneous variants of S194-2 myeloma cells that have lost IgA secretion.
- To investigate the frequency and biochemical properties of these non-secreting variants.
Main Methods:
- Cloning S194-2 cells on soft agar and screening for loss of secreted immunoglobulin.
- Utilizing DNP-ferritin conjugates to specifically detect IgA secretion via precipitation.
- Biochemical analysis of isolated variants.
Main Results:
- Spontaneous variants with reduced or undetectable IgA secretion were isolated at a frequency of 2.7 X 10(-5) per cell per generation.
- No variants were found with altered DNP binding sites or those secreting only immunoglobulin subunits.
- An enhanced S194 culture secreting 5-6 times more IgA was developed through selection.
Conclusions:
- Spontaneous loss of IgA secretion occurs in S194-2 myeloma cells at a significant frequency.
- The isolated variants primarily exhibit a complete or near-complete loss of IgA secretion, without specific alterations in the DNP binding site or subunit secretion.