SummaryA set of 13 hybridoma antibodies to type 1 poliovirus has been studied with regard to neutralization and bindi_ng to N antigen, H antigen and capsid proteins. Two hybridomas produced heterogeneous antibodies, even after repeated attempts to reelone them. ,A set of thirteen mouse hybridoma antibodies to type 1 poliovirus was developed. The antibodies were studied with regard to a) neutralization of type 1, 2 and 3 poliovirus; b) reaction with N antigen, i.e. native, mature virions; c) reaction with H antigen, i.e., heat-denatured virions (6); d) reaction with individual capsid polypeptides prepared by detergent disruption of the virions.Two subsets of antibodies are tentatively distinguished: a) neutralizing monoelones recognizing N antigen only, and b) non-neutralizing monoctones, which recognized H antigen; some of these were also able to bind to the capsid polypeptides. The case of two hybridomas which produced antibodies sharing both sets of properties will be discussed.To prepare N antigen, type 1 (strain I a/S 3) poliovirus was grown and purified as described (10), and freed of empty eapsids by sucrose gradient eentrifugation and collection of the virions as 160 S particles. One microgram of this antigen emulsified in Freund's complete adjuvant was used for the initial immunization of BALB/c mice by intraperitoneal injection. A single booster of 1 ~zg N antigen was given intravenously after 1 month, and the animals were sacrificed 4 to 6 days later. The spleen cells were fused with SP2/0 mouse myeloma cells in the ratio of 10:1. Fusion and hybrid selection were done by conventional methods using PEG 1500 and ttAT medium (3).tIybmdomas secreting polio-specific antibodies were screened for by a doublesandwich ELISA method. The capture layer was formed by coating polystyrene 0304-8608/82/0074/0325/$ 01.20

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