Surface proteins and glycoproteins of ex- sheathed L3 (XL3), L4 and adults were identified by labelling live worms with NHS-SS-biotin which labels primary amine groups and biotin hydrazide (BH) which introduces biotin into sugar residues. After labelling and extensive washing, worms were homogenised in low-osmotic-strength buffer (TBS) and subjected to sequential extractions with SDS and SDS + 2-Me containing buffers. Labelled proteins released from the worms by these procedures were analysed by SDS-PAGE followed by Western blotting and probing with AP-streptavidin. Western blot analysis of biotinylated cuticular proteins from parasitic stages of O. circumcincta revealed stage specific differences in the labelling patterns of the cuticular proteins and glycoproteins. Developmental differences in cuticular components were apparent in the variety of MW of the components seen in Western blots although some components identified in Western blots appeared to be conserved (78 and 45 kDa). Two to six polypeptides were recognised in XL3 surface extracts by serum from sheep vaccinated with adult nematode somatic proteins. The surface of fourth stage larvae showed less cross-reactivity. Interestingly, the molecular weights of polypeptides recognised by anti-adult serum in streptavidin affinity isolated XL3 cuticular proteins were different from those recognised in adult cuticular proteins, which suggested that the cross-reacting epitopes were located on different proteins.
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