Selection of large diversities of antiidiotypic antibody fragments by phage display.

摘要: Antiidiotypic antibodies (Ab2) are needed as tools for a better understanding of molecular mimicry and the immunological network, many potential applications in biomedical pharmaceutical field. mimicking carbohydrate or conformational epitopes (Ab2beta) considerable interest surrogate immunogens cancer vaccination. However, it has so far been difficult tedious to produce Ab2s given antigen. Here we describe fast reliable technique generating large diversities antiidiotypic single chain antibody fragments from non-immunized phagemid libraries using phage display. Key elements specific elution with original antigen followed by trypsin treatment eluted phages combination protease sensitive helperphage KM13. This novel method was compared various conventional selection methods, including, without treatment, glycine at pH 2.2 only. The results clearly show that is superior other enabling first time generation variety after only two three rounds selection, thereby maintaining maximum diversity. We obtained 28 88 antiidiotypes out 96 tested clones diversity 55-90 %. achieved (di-, tetrasaccharides) one protein epitope naive their corresponding monoclonal Ab1. nature selected scFv-phages verified ELISA immunocytochemistry inhibition experiments.