Studies involving the specific and non-specific surface recognition of biotin on biotinylated LB lipid monolayers by streptavidin and avidin conjugated phycoerythrin are presented. Both streptavidin and avidin conjugates were injected under the monolayer and found to preferentially adsorb to the biotinylated monolayers at the air-water interface. Pressure-area isotherms displayed a biotin-streptavidin/avidin complex dependent increase in surface pressure at expanded areas indicating protein adsorption. The binding of protein was confirmed by transferring the monolayer films to solid supports and measuring the characteristic intense phycoerythrin fluorescence at 576 nm. The effect of protein charge, monolayer packing density and structure, and activation of the tetramer proteins towards specific and non-specific binding are discussed. These results suggest a novel and general methodology for the two-dimensional ordering of protein monolayers with potential bioelectronic, optical and protein structure research applications.