Anti-survivin single-domain antibodies derived from an artificial library including three synthetic random regions by in vitro selection using cDNA display
| Autor: | Naoto Nemoto, Takeru Suzuki, Yoshihisa Hagihara, Shinnosuke Kimura, Yoko Akazawa-Ogawa, Yuki Mochizuki |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
DNA Complementary Survivin Biophysics Computational biology Inhibitor of apoptosis Biochemistry 03 medical and health sciences Complementary DNA Animals Amino Acid Sequence Molecular Biology Gene Library 030102 biochemistry & molecular biology Heavy-chain antibody biology Brevibacillus Chemistry Cell Biology Single-Domain Antibodies Surface Plasmon Resonance In vitro 030104 developmental biology Single-domain antibody biology.protein Target protein Antibody Protein Binding |
| Zdroj: | Biochemical and biophysical research communications. 503(3) |
| ISSN: | 1090-2104 |
| Popis: | Single-domain antibodies (variable domain of the heavy chain of a heavy chain antibody; VHH) are promising reagents for therapeutics and diagnostics because of their stability, cost-effective production and material workability as a small antibody. Currently, general acquisition of a VHH using immunization of camelids is inconvenient from the standpoint of animal protection, cost and the process is time-consuming. Thus, a straightforward and efficient method for screening VHHs against a target molecule is required. In this study, we examined whether in vitro selection of a VHH against a target protein could be performed by a cDNA display method with an artificial VHH library that had the three complementarity-determining regions (CDRs) randomized by chemical synthesis. The results revealed that a particular VHH against survivin, which is a member of the inhibitor of apoptosis family, was selected with affinity in the range of 10−7 to 10−8 M. The in vitro selection of a VHH using cDNA display with an artificial synthesized library without animal immunization was shown to be effective for rapid and inexpensive screening of VHHs against a target protein. |
| Databáze: | OpenAIRE |
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