8IPC
The recombinant NZ-1 Fab complexed with the PDZ tandem fragment of A. aeolicus S2P homolog with the PA14 tag inserted between the residues 181 and 184
Summary for 8IPC
| Entry DOI | 10.2210/pdb8ipc/pdb |
| Descriptor | The recombinantly-expressed heavy chain of the monoclonal antibody NZ-1, The recombinantly-expressed light chain of the monoclonal antibody NZ-1, Putative zinc metalloprotease aq_1964, ... (4 entities in total) |
| Functional Keywords | pdz domain fab, hydrolase, immune system-hydrolase complex, immune system/hydrolase |
| Biological source | Rattus norvegicus More |
| Total number of polymer chains | 3 |
| Total formula weight | 68036.92 |
| Authors | Adachi, Y.,Nogi, T. (deposition date: 2023-03-14, release date: 2024-01-31, Last modification date: 2024-11-20) |
| Primary citation | Adachi, Y.,Kaneko, M.K.,Kato, Y.,Nogi, T. Recombinant production of antibody antigen-binding fragments with an N-terminal human growth hormone tag in mammalian cells. Protein Expr.Purif., 208-209:106289-106289, 2023 Cited by PubMed Abstract: Antigen-binding fragments (Fabs) of antibodies are both key biopharmaceuticals and valuable tools for basic life science. To streamline the production of diverse Fabs by capitalizing on standard and highly optimized protein production protocols, we here explore a method to prepare recombinant Fabs as secreted fusion proteins with an N-terminal human growth hormone domain and an octa-histidine tag. These tagged Fabs can be purified with standard immobilized metal chelate affinity chromatography. We first demonstrated Fab overproduction using the rat monoclonal antibody NZ-1. Optimization of linker residues enabled the complete removal of the tags by TEV protease, leaving only two additional residues at the N-terminus of the heavy chain. We purified NZ-1 Fab at ∼4 μg/mL of culture supernatant and further confirmed that the NZ-1 Fab from the fusion protein maintained its native fold and binding affinity for target cell-surface antigens. We also showed that several other Fabs of mouse IgGs, the major subclass in mice, could be produced with the same procedure. Our preparation method can provide greater flexibility in functional and structural modifications of target Fabs because specialized purification techniques are not necessary. PubMed: 37160213DOI: 10.1016/j.pep.2023.106289 PDB entries with the same primary citation |
| Experimental method | X-RAY DIFFRACTION (2.2 Å) |
Structure validation
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