Cell cycle dependency of monoclonal antibody production in asynchronous serum-free hybridoma cultures

Abstract

The cell cycle kinetics of F3(B6) mouse hybridoma was examined by immunocytochemical staining of bromodeoxyuridine incorporated into the DNA of exponentially growing cells in three different cultures: one supplemented with 10% fetal bovine serum and two adapted to serum-free media, TABIES and BITES. The serum-free cultures, particularly the BITES, had longer cycling times and higher specific antibody production rate. Both observations were correlated to the prolongation of the G1 phase traverse time and substantiated with a starvation blocking experiment.

Cell Culture Engineering VI

“…This is in agreement with a number of studies reporting that synthesis and secretion of immunoglobulins are maximal in late Ol/early S phase (Byars and Kidson, 1970;Oaratun-tjeldsto et al, 1976;Suzuki and Ollis, 1989;Ramirez and Mutharasan, 1990;Richieri et al, 1991;Cazzador and Mariani, 1993). In a previous study we have shown that treatment of D5 hybridoma cells with OptiMAb™ had also resulted in increased qMAb and final MAb concentration, associated with prolongation of the 01 phase .…”

Apoptosis-resistant E1B-19K-expressing NS/0 myeloma cells exhibit increased viability and chimeric antibody productivity under perfusion culture conditions

“…This has been observed for cells cultivated in batch (Ramirez and Mutharasan, 1990), fed-batch (de Tremblay et al, 1993;Robinson et al, 1994), chemostat (Boraston et al, 1984), and perfusion modes (Al-Rubeai et al, 1992;Banik and Heath, 1995;Batt et al, 1990;de la Broise et al, 1991;Johnson et al, 1996;Hansen et al, 1993;Park and Ryu, 1994;Tokashiki and Takamatsu, 1993;Trampler et al, 1994), as well as in cells treated with cell-cycle blocking agents (Al-Rubeai and Emery, 1990;Jayme, 1991;Mercille and Massie, 1998;Oh et al, 1995;Øyaas et al, 1994a,b;Reddy and Miller, 1994;Suzuki and Ollis, 1990;Takahachi et al, 1994). Indeed, specific antibody production has been shown to be cell cycle dependent and optimum in the G 1 and/or G 2 /M phase (Cazzador and Mariani, 1993;Kromenaker and Srienc, 1991;Linardos et al, 1992;Richieri et al, 1991;Suzuki and Ollis, 1989). However, conditions leading to retardation of lymphoid cell growth are also a potent inducer of apoptosis in a variety of cell lines.…”

“…Indeed, many mammalian cell lines cultivated in perfusion exhibit an increase in specific MAb productivity under such slow growth conditions (Al-Rubeai et al, 1992;Banik and Heath, 1995;Batt et al, 1990;de la Broise et al, 1991de la Broise et al, , 1992Johnson et al, 1996;Hansen et al, 1993;Mercille et al, 1994a-c;Shi et al, 1993;Tokashiki and Takamatsu, 1993;Trampler et al, 1994). The q MAb has been shown to be cell cycle dependent and optimum in the G 1 (Byars and Kidson, 1970;Cazzador and Mariani, 1993;Garatun-Tjeldsto et al, 1976;Linardos et al, 1992;Mercille and Massie, 1998;Ramirez and Mutharasan, 1990;Richieri et al, 1991;Suzuki and Ollis, 1989) or G 2 /M phase (Cherlet et al, 1995;Kromenaker and Srienc, 1991;Mercille and Massie, 1998;Watanabe et al, 1973). The increase in the perfusion to batch q MAb ratio can therefore be explained by an increase in the percentage of G 1 cells in perfusion from 40 ± 5% for NS/0 control cells to 57 ± 6% for E1B-19K cells.…”

Cell cycle dependency of rice α-amylase production in a recombinant yeast

“…It was reported that antibody synthesis and secretion rate were regulated during the cell cycle (Al-Rubeai and Emery, 1990;Cherlet et al, 1995;Kromenaker and Srienc, 1991;Martens et al, 1993;Richieri et al, 1991). Assays for tissue-like plasminogen activator (tPA) and urokinase-like plasminogen activator (uPA) produced by a rat adenocarcinoma cell line, PA-III, revealed an increased appearance of both enzymes shortly after the S phase (Scott et al, 1987).…”