Abstract
BackgroundConsistent progress in the development of bacteriophage lambda display platform as an alternative to filamentous phage display system was achieved in the recent years. The lambda phage has been engineered to display efficiently multiple copies of peptides or even large protein domains providing a powerful tool for screening libraries of peptides, proteins and cDNA.ResultsIn the present work we describe an original method for dual display of large proteins on the surface of lambda particles. An anti-CEA single-chain antibody fragment and green fluorescent protein or alkaline phosphatase were simultaneously displayed by engineering both gpD and gpV lambda proteins.ConclusionsHere we show that such modified phage particles can be used for the detection of target molecules in vitro and in vivo. Dual expression of functional moieties on the surface of the lambda phage might open the way to generation of a new class of diagnostic and therapeutic targeted nanoparticles.
Highlights
Consistent progress in the development of bacteriophage lambda display platform as an alternative to filamentous phage display system was achieved in the recent years
The capacity of a foreign protein to be secreted across the membrane depends on its amino acid composition [4,6,7,8], filamentous phage-based libraries only display those recombinant proteins able to pass through the inner bacterial membrane still maintaining their correct folding in the oxidizing environment of the periplasmic space
An anti-Carcino embryonic antigen (CEA) single-chain antibody fragment and green fluorescent protein or alkaline phosphatase were simultaneously displayed by engineering both gpD and gpV lambda proteins
Summary
Consistent progress in the development of bacteriophage lambda display platform as an alternative to filamentous phage display system was achieved in the recent years. A very low number of larger polypeptides can be displayed by fusion to the tolerant minor protein pIII, which is only present in five copies at one end of the phage filament In this latter case the avidity of the recombinant phage for its ligand is dramatically reduced, seriously limiting the selection efficiency of ligands for receptors only available at low concentration or present in complex mixtures (as in the case of biological fluids, such as serum). Another factor that can influence the exposition of the foreign protein on the phage surface is connected with the viral life cycle. The capacity of a foreign protein to be secreted across the membrane depends on its amino acid composition [4,6,7,8], filamentous phage-based libraries only display those recombinant proteins able to pass through the inner bacterial membrane still maintaining their correct folding in the oxidizing environment of the periplasmic space
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