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A Sensitive, Single-Radial Diffusion Autoradiographic Zone Size Enhancement Technique for the Assay of Influenza Haemagglutinin

Division of Viral Products, National Institute for Biological Standards and Control, Holly Hill, Hampstead, London NW3 6RB, U.K.

A novel radial-diffusion technique in agarose gels is described which is applicable to the assay of small concentrations of influenza haemagglutinin (HA) antigen. The method is based on single-radial-diffusion zone size enhancement (ZE), using autoradiography to demonstrate antigen-antibody reactions. The ZE technique is capable of reliably assaying concentrations of haemagglutinin as low as 0.1 µg/ml, which represents a 20- to 40-fold increase in sensitivity over conventional single-radial-diffusion techniques. The ZE response is dependent upon the antigenic identity of the ¹²⁵I-labelled ‘marker’ HA and the HA antigen being assayed. Haemagglutinins showing variation within a subtype (e.g. H3) show reduced ZE responses and thus the method has potential value for distinguishing between closely related HA antigens.

Received 12 September 1979; accepted 31 October 1979.