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.

Immunological Evidence for the Existence of a Carrier Protein for Sucrose Transport in Tonoplast Vesicles from Red Beet (Beta vulgaris L.) Root Storage Tissue

“…Following fusion, cells were plated into 24-well Nunc plates and grown in RPMI medium supplemented with 10% fetal calf serum (Flow Laboratories), antibiotics, 2 m~ Glu, 100 p~ hypoxanthine, 100 p~ azaserine as selective medium (Zagury et al, 1979).…”

Mathematical models of cytotoxic T‐lymphocyte killing

“…While in the chromium release assay one can measure the number of killed target cells on a population level, subsequent advances in experimental techniques allowed the observation of CTL killing at the level of single cells 17 , 18 , 19 , 20 . The new experimental technique (referred to as the CMC assay in agarose) involved the isolation of so‐called conjugates – multicellular complexes in which CTL and target cells were bound to each other.…”

Influence of Dehydrocholic and Cholic Acids on the Biliary Secretion of Anionic Polypeptide Fraction, the Major Apoprotein of the Biliary Lipoprotein Complex

“…Fusions were performed as described by Nowinsky et al (19), using SP2-0 Ag 14 myeloma cells (20). The selection medium contained azaserine and hypoxanthine (21).…”