Production of rat monoclonal antibody from rat x mouse hybridoma cell lines using microencapsulation technology.
The cell growth and monoclonal antibody production characteristics of two rat x mouse heterohybridoma cell lines, designated 187.1 and M1/9.3, were investigated using a biocompatible microencapsulation technology. Both cell lines, derived from the fusion of immunized rat spleen cells with either the NS1 or X63Ag8.653 myeloma cell lines, were found to reach a maximum intracapsular cell density of 1.3 to 1.5 X 10(7) cells/ml during a 27-d culture period. During this period, rat monoclonal antibody accumulated in the intracapsular space of both cultures to a final concentration of 2.0 to 2.8 mg/ml. Comparison of the concentration of rat monoclonal antibody in the extracapsular vs. the intracapsular space of both cultures indicated that significantly less than 1% of the antibody produced by the encapsulated hybridoma cells was capable of transiting the microcapsule membrane during the culture period. Due to the partition of the rat monoclonal antibody within the intracapsular space, the initial purity of the antibody harvested from 21-d microcapsule cultures of 187.1 and M1/9.3 cells was approximately 48 and 75% by weight, respectively. Analysis of the intracapsular protein by sodium dodecyl sulfoxide gel electrophoresis at different times during the culture period demonstrated that the principal contaminant associated with the unpurified antibody was bovine serum albumin.
Gilligan
K J
KJ
Damon Biotech, Inc., Needham Heights, Massachusetts 02194.
Littlefield
S
S
Jarvis
A P
AP
D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000302:isolation & purification; D002455:Cell Division; D004591:Electrophoresis, Polyacrylamide Gel; D006825:Hybridomas / Q000166:cytology / Q000276:immunology*; D007118:Immunoassay; D051379:Mice; D011506:Proteins / Q000032:analysis; D051381:Rats
D000911:Antibodies, Monoclonal; D011506:Proteins