Anti-peroxidase antibody-secreting hybrid lines. I. Identification, cloning and cell characterization

Abstract

Anti-peroxidase antibody (Ab)-secreting hybrids have been produced by fusion of peroxidase (PO)-immunized mouse lymph node cells and immunoglobulin (Ig)-secreting P3-X63-Ag8 (X63) myeloma cells. Identification of Ab-secreting hybrids can be performed as early as day 5 after cell fusion by the hemolytic plaque assay. Immediately after identification, hybrids were directly isolated, by means of a micropipette, into Terasaki microchambers containing nutrient medium and a thymocyte filler layer. The yield of secreting hybrids is improved by using this procedure. All the cells of the PO 772 C2 clone show the same ultrastructural pattern and immunocytological properties; they are proplasmocytes, as are the parental X63 cells; they present intracisternae Ab and show no Ig or Fc receptors at the cell surface. Over 90% of viable PO 772 C2 cells form specific plaques. Isoelectric focusing and sodium dodecyl sulfate polyacrylamide gel electrophoresis show that the cells of this clone secrete Ab; the secreted Ig are formed with chi and gamma 1 chains from the parental X63 cells and specific L and H chains from the lymphoid parent. These biological investigations demonstrate the relative stability of the PO 772 C2 clone secreting anti-peroxidase antibody.