The potential of flow cytometric analysis for the characterization of hybridoma cells in suspension cultures.
Flow cytometric (FC) analysis was applied to determine changes at cellular level during the cultivation of hybridoma cell line MN12 in a suspension batch culture. The relative cell size, cytoplasmic and membrane IgG content and the viability were monitored. Besides, the specificity of the cytoplasmic and membrane IgG was ascertained by means of a synthetic peptide containing the antigenic epitope recognized by the antibody. Cell size was found to increase during the exponential growth phase. The viability as determined by FC follows a similar pattern with the viability data obtained by the conventional trypan blue exclusion test. The relative cytoplasmic and membrane IgG contents were high during the exponential growth and low during stationary phase. Measurement of cell cycle distribution and the antibody content in the culture fluid, indicated that the major part of the cytoplasmic IgG is secreted by cells in the G1-phase. It is concluded that flow cytometry is a useful tool to characterize hybridoma cell lines in a suspension batch culture.
Department of Inactivated Viral Vaccines, National Institute of Public Health and Environmental Protection (RIVM), Bilthoven, The Netherlands.
J M
JM
Coco-Martin
J W
JW
Oberink
T A
TA
van der Velden-de Groot
E C
EC
Beuvery
D000595:Amino Acid Sequence; D000911:Antibodies, Monoclonal / Q000096:biosynthesis; D000918:Antibody Specificity; D002453:Cell Cycle; D002455:Cell Division; D002462:Cell Membrane / Q000737:chemistry; D002470:Cell Survival; D003593:Cytoplasm / Q000737:chemistry; D005069:Evaluation Studies as Topic; D005434:Flow Cytometry; D006825:Hybridomas / Q000737:chemistry / Q000166:cytology* / Q000378:metabolism; D007074:Immunoglobulin G / Q000032:analysis; D007700:Kinetics; D008969:Molecular Sequence Data
D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G