ArticleBiological procedures online2017
Efficient and inexpensive transient expression of multispecific multivalent antibodies in Expi293 cells.
Article in Biological procedures online, 2017. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 68 papers.
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68 citing papers in PubMed, 116 citations in OpenAlex.
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- Quality by design for transient RBD-Fc fusion protein production in Chinese hamster ovary cells.Biotechnology reports (Amsterdam, Netherlands) · 2025Article
- Enhancing therapeutic antibody profiling: orthogonal strategies for stability and quality assessment.Frontiers in pharmacology · 2025Article
- Updated transient gene expression protocol in Expi293F cells using PEI.Frontiers in bioengineering and biotechnology · 2025Article
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- A shorter linker in the bispecific antibody RmAb158-scFv8D3 improves TfR-mediated blood-brain barrier transcytosis in vitro.Scientific reports · 2024Article
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- Mapping the immunopeptidome of seven SARS-CoV-2 antigens across common HLA haplotypes.Nature communications · 2024Article
- Episomal Vectors for Stable Production of Recombinant Proteins and Engineered Antibodies.Antibodies (Basel, Switzerland) · 2024Article
- Altered amyloid-β structure markedly reduces gliosis in the brain of mice harboring the Uppsala APP deletion.Acta neuropathologica communications · 2024Article
- Extracellular carriers control lipid-dependent secretion, delivery, and activity of WNT morphogens.Developmental cell · 2024Article
- A charged tail on anti-α-Synuclein antibodies does not enhance their affinity to α-Synuclein fibrils.PloS one · 2024Article
- Indium-111 radiolabelling of a brain-penetrant Aβ antibody for SPECT imaging.Upsala journal of medical sciences · 2024Article
8 more citing papers are in PubMed but not listed here.
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Authors and funding
5 authors at 1 institution in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
backgroundImmunotherapy is a very fast expanding field within drug discovery and, hence, rapid and inexpensive expression of antibodies would be extremely valuable. Antibodies are, however, difficult to express. Multifunctional antibodies with additional binding domains further complicate the expression. Only few protocols describe the production of tetravalent bispecific antibodies and all with limited expression levels..
methodsHere, we describe a protocol that can produce functional tetravalent, bispecific antibodies at around 22 mg protein/l to a low cost. The expression system is based on the Expi293 cells, which have been adapted to grow in denser cultures than HEK293 cells and gives higher expression yields. The new protocol transfects the Expi293 cells with PEI (which has a negligible cost).
resultsThe protocol has been used to generate multiple variants of tetra- and hexavalent bispecific antibodies with yields of around 22 mg protein/l within 10 days. All materials are commercially available and the implementation of the protocol is inexpensive and straightforward. The bispecific antibodies generated in our lab were capable of binding to all antigens with similar affinity as the original antibody. Two of the bispecific antibodies have also been used in transgenic mice as positron emission tomography (PET) ligands to successfully detect amyloid-beta (Aβ) aggregates
conclusionsThis protocol is the first describing transfection of the human Expi293 cells with PEI. It can be used to generate functional multi-specific antibodies in high amounts. The use of biological drugs, and in particular multispecific antibodies, is rapidly increasing, hence improved protocols such as the one presented here are highly valuable.
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