A1 Vertaisarvioitu alkuperäisartikkeli tieteellisessä lehdessä
Two ScFv antibody libraries derived from identical VLVH framework with different binding site designs display distinct binding profiles
Tekijät: Huovinen T, Syrjanpää M, Sanmark H, Brockmann EC, Azhayev A, Wang Q, Vehniäinen M, Lamminmäki U
Julkaisuvuosi: 2013
Journal: Protein Engineering, Design and Selection
Numero sarjassa: 10
Vuosikerta: 26
Numero: 10
Aloitussivu: 683
Lopetussivu: 693
Sivujen määrä: 11
ISSN: 1741-0126
DOI: https://doi.org/10.1093/protein/gzt037
Tiivistelmä
In directed evolution experiments, a single randomization scheme of an antibody gene does not provide optimal diversity for recognition of all sizes of antigens. In this study, we have expanded the recognition potential of our universal library, termed ScFvP, with a second distinct diversification scheme.In the second library, termed ScFvM, diversity was designed closer to the center of the antigen binding site in the same antibody framework as earlier. Also, the CDR-H3 loop structures were redesigned to be shorter, 512 aa and mostly without the canonical salt bridge between Arg106(H) and Asp116(H) to increase the flexibility of the loop and to allow more space in the center of the paratope for binding smaller targets. Antibodies were selected from the two libraries against various antigens separately and as a mixture. The origin and characteristics of the retrieved antibodies indicate that complementary diversity results in complementary functionality widening the spectrum of targets amenable for selection.
In directed evolution experiments, a single randomization scheme of an antibody gene does not provide optimal diversity for recognition of all sizes of antigens. In this study, we have expanded the recognition potential of our universal library, termed ScFvP, with a second distinct diversification scheme.In the second library, termed ScFvM, diversity was designed closer to the center of the antigen binding site in the same antibody framework as earlier. Also, the CDR-H3 loop structures were redesigned to be shorter, 512 aa and mostly without the canonical salt bridge between Arg106(H) and Asp116(H) to increase the flexibility of the loop and to allow more space in the center of the paratope for binding smaller targets. Antibodies were selected from the two libraries against various antigens separately and as a mixture. The origin and characteristics of the retrieved antibodies indicate that complementary diversity results in complementary functionality widening the spectrum of targets amenable for selection.
Research Areas
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