Production of a murine monoclonal antibody with anti-hiv-1 neutralizing properties

✍ Scribed by Jorma Hinkula; Gustav Gilljam; Britta Wahren

• Publisher: John Wiley and Sons
• Year: 1991
• Tongue: English
• Weight: 608 KB
• Volume: 5
• Category: Article
• ISSN: 0887-8013
• DOI: 10.1002/jcla.1860050607

✦ Synopsis

A mouse monoclonal hybridoma cell line producing IgG 1 k to human immunodeficiency virus (HIV-1) gp120 envelope protein was cultured in several systems. A small-scale flask culture was essential for characterizing the culture variables of the hybridoma. A dialysis tubing culture appeared to be an excellent alternative to in vivo cultures of ascitic fluid, and gave high mouse monoclonal antibody (Mab) concentrations. Two continuous culture systems were both very effective in producing large amounts of Mabs. The hollow fiber system has the advantage of giving a concentrated product in the harvest. The ceramic core system, on the other hand, allows excellent monitoring of the cellular growth and production phases and gave the highest HIV antigen reactivity/pg of the produced IgG.

Twelve grams of HIV-1 neutralizing Mabs were produced. The Mab was purified with a yield of 61%. The neutralizing capacity of the Mab was studied in vitro and shown to be excellent with 50% neutralizing titers using 5 ng Mab. The biological half-life of the Mab given intraveneously to an HIV-infected individual was shown to be around 30 h.