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Reliable cloning of functional antibody variable domains from hybridomas and spleen cell repertoires employing a reengineered phage display system.
Fully synthetic human combinatorial antibody libraries (HuCAL) based on modular consensus frameworks and CDRs randomized with trinucleotides.
The small number of 49 master genes will allow future improvements to be incorporated quickly, and the separation of the frameworks may help in analyzing why nature has evolved these distinct subfamilies of antibody germline genes.
Designing repeat proteins: well-expressed, soluble and stable proteins from combinatorial libraries of consensus ankyrin repeat proteins.
Assembly of a functional immunoglobulin Fv fragment in Escherichia coli.
An expression system was developed that allows the production of a completely functional antigen-binding fragment of an antibody in Escherichia coli, and experiments showed that the affinity constant of the Fv fragment is identical to that of the native antibody McPC603, that there is one binding site for phosphorylcholine in the FV fragment, and that there are no inactive protein in the preparation.
In vitro selection and evolution of functional proteins by using ribosome display.
Libraries of native folded proteins can now be screened and made to evolve in a cell-free system without any transformation or constraints imposed by the host cell.
Folding and unfolding mechanism of highly stable full-consensus ankyrin repeat proteins.
Stability engineering of antibody single-chain Fv fragments.
Progress in rational and evolutionary engineering methods, the structural implications of these results, as well as some examples where stability engineering has been successfully applied are summarized.
Crystal Structure of the Dimeric C-terminal Domain of TonB Reveals a Novel Fold*
- Changsoo Chang, Alexandre Mooser, A. Plückthun, A. Wlodawer
- ChemistryThe Journal of Biological Chemistry
- 20 July 2001
The crystal structure of the C-terminal domain of TonB from Escherichia coli has been solved by multiwavelength anomalous diffraction and refined at 1.55-Å resolution, providing the first evidence that this region ofTonB dimerizes.
Induction and Exhaustion of Lymphocytic Choriomeningitis Virus–specific Cytotoxic T Lymphocytes Visualized Using Soluble Tetrameric Major Histocompatibility Complex Class I–Peptide Complexes
- A. Gallimore, A. Glithero, R. Zinkernagel
- Biology, MedicineThe Journal of experimental medicine
- 4 May 1998
The results of the study show that the outcome of infection by LCMV is determined by antigen load alone, and deletion of virus-specific CTLs in the presence of excessive antigen is preceded by TCR downregulation and is dependent upon perforin.
In vitro generated antibodies specific for telomeric guanine-quadruplex DNA react with Stylonychia lemnae macronuclei
- C. Schaffitzel, I. Berger, J. Postberg, J. Hanes, H. Lipps, A. Plückthun
- Biology, ChemistryProceedings of the National Academy of Sciences…
- 3 July 2001
Experimental evidence is provided that the telomeres of Stylonychia macronuclei adopt in vivo a guanine-quadruplex structure, indicating that this structure may have an important role for telomere functioning.