The Ly-25.1 specificity: Definition with a monoclonal antibody

✍ Scribed by P. Mark Hogarth; Aveline Rigby; Vivien R. Sutton; Ian F. C. McKenzie; J. Hilgers

• Book ID: 104741173
• Publisher: Springer-Verlag
• Year: 1984
• Tongue: English
• Weight: 220 KB
• Volume: 19
• Category: Article
• ISSN: 0093-7711
• DOI: 10.1007/bf00364478

✦ Synopsis

To study individual cell surface molecules of murine lymphocytes, we are producing monoclonal antibodies defining both new and existing alloantigenic determinants. In this context, we now describe a new alloantigenic system -Ly-25 -defined by monoclonal antibody 5069-1.1 detecting the Ly-25.1 specificity. The specificity is found only on hemopoietic cells.

To produce this antibody, (C3H x DBA/2)F1 mice were immunized six times with CE/J thymocytes over a period of 3 months with a final intravenous injection of 107 thymocytes. Three days after the final injection, spleen cells from the immunized mice were fused with the NS-1 myeloma (Hogarth et al. 1980). Hybridomas secreting specific antibodies were detected by screening culture supernatants using a rosetting method with sheep anti-mouse Ig-coated erythrocytes (SAMIg-rosetting) (Parish and McKenzie 1978) and CE thymocytes as target cells. Cell line 5069-1.l (described herein) was cloned twice and was found to secrete an IgG2a antibody that was not cytotoxic with rabbit complement and did not bind staphylococcal protein A.