Selection and purification of L1 scFvs from the Tomlinson I library.

<p>(<b>A)</b> Binding of monoclonal phages to substrate-coated L1/ecd was determined by ELISA. Phages (1×10⁹ cfu) from the third selection round were incubated with L1/ecd. Substrate-coated BSA was used as negative control. Bound phages were detected with anti-M13 antibody conjugated to HRP. (<b>B</b>) Expression and purification of scFvs. Lane 1, bacterial supernatant containing scFvs; lane 2, column flow-through; lane 3, affinity chromatography purified scFv I27 (indicated by arrow) following column elution with 100 nM imidazole. Molecular weight markers are indicated at the left margin. (<b>C</b>) Amino acids sequences of complementarity determination regions 2 and 3 on heavy and light chains of L1 scFvs. Different amino acids are underlined.