The Hybridoma Facility of the Laboratory of Immunoregulation has collaborated with numerous laboratories in FDA and NIH in the production and characterization of monoclonal antibodies (MAbs). During this reporting period the facility has completed and assisted in 13 hybridoma fusions. These fusions required screening by ELISA, immuno-precipitation, gel shift, FACS analysis, Western blot, dot blot, and in vivo bioactivity assay of approximately 11,022 wells of tissue culture fluid. As a result 947 lines and 62 clones (cloning from two fusions is in progress and not included in this number) have been selected for further study and subsequently grown up and frozen. Ascites fluid has been produced to 64 cell lines, totaling 1086 ml volume. This years work compares to the six year averages (1988-1993) as follows (1994/6 year average): fusions, 13/15; lines selected per fusion, 77/16; lines screened per fusion, 848/367 and ascites per fusion, 5/2.9. Procedures have been initiated for an additional 7 fusions to be completed during the next reporting period. One of the fusions was also used for the complete training in the technology of a visiting scientist. The facility has also conducted ancillary procedures from 17 previously completed fusions. These include ascites production to 21 lines yielding 959 ml of fluid containing MAbs, production of 1525 ml of MAb containing culture fluid, production of polyclonal sera in mice, freezing new stocks of cell lines and aliquoted ascites fluids and shipping cell lines and fluids. This year a total of 635 mice were used for immunization or ascites production. Four different growth supplements were evaluated during 3 fusion procedures, a more effective one selected and put into regular use at the facility. The standard operating procedures manual was edited and provided to 4 investigators. Two internal and one external manuscripts for publication in scientific journals were reviewed. Three copies of a computer program written by the facility manager to log data on cells stored in liquid nitrogen freezers were distributed externally free of cost to bring the total distribution to 156 copies. The facility operations were twice moved out of the laboratory due to renovations of HVAC and cabinetry into temporary space, and then returned. The core Reviewer Training was completed and certified, including modules I, II, IV, VII and VIII as well as extra module VI, a total of 28 1/2 hours. The core Inspector Training was also completed and certified including modules I, II, III and extra module IV, a total of 18 hours. Other training included personal computer topics, radiation safety refresher training, user fee information, sexual harassment prevention training, and sterilization tunnel validation and clean room design.
