Molecular Biological Analysis of Perfusion Culture of Hybridoma and Effective Production of Monoclonal Antibody Using Interleukin-6

Abstract

Hybridoma 2E3 cells produced two to three times as much antibody per cell in perfusion culture as did cells growing exponentially in batch culture. The proportion of γ₁ mRNA to total RNA of the cells in perfusion culture was 1.7 times as much as that of the cells growing exponentially in batch culture. The growth rate of the cells in perfusion culture, determined by thymidine incorporation rate into DNA of the cells, was about one-third that of cells growing exponentially in batch culture. Cell cycle analysis using flow cytometry indicated that the cells which were growth-suppressed in the perfusion culture accumulated in G₁ phase of the cell cycle. Concentrations of ammonium ion, glucose and lactate in the supernatant of the perfusion culture were almost equal to those of the late batch culture. Interleukin-6 enhanced the specific antibody production rate of the cell in perfusion culture.