Change search
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf
Efficient and inexpensive transient expression of multispecific multivalent antibodies in Expi293 cells
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Public Health and Caring Sciences, Geriatrics. (Molekylär geriatrik/ Rudbecklaboratoriet)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Public Health and Caring Sciences, Geriatrics. (Molekylär geriatrik/ Rudbecklaboratoriet)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Public Health and Caring Sciences, Geriatrics. (Molekylär geriatrik/ Rudbecklaboratoriet)
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Public Health and Caring Sciences, Geriatrics. (Molekylär geriatrik/ Rudbecklaboratoriet)
Show others and affiliations
2017 (English)In: Biological Procedures Online, ISSN 1480-9222, E-ISSN 1480-9222, Vol. 19, article id 11Article in journal (Refereed) Published
Abstract [en]

Background: Immunotherapy 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.

Methods: Here, 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).

Results: The 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 (A beta) aggregates in vivo.

Conclusions: This 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.

Place, publisher, year, edition, pages
2017. Vol. 19, article id 11
Keyword [en]
Bispecific antibodies, Expi293, HEK293, PEI, Transient protein expression
National Category
Biochemistry and Molecular Biology
Identifiers
URN: urn:nbn:se:uu:diva-335186DOI: 10.1186/s12575-017-0060-7ISI: 000410951400001PubMedID: 000410951400001OAI: oai:DiVA.org:uu-335186DiVA, id: diva2:1163939
Available from: 2017-12-08 Created: 2017-12-08 Last updated: 2017-12-08Bibliographically approved

Open Access in DiVA

fulltext(913 kB)8 downloads
File information
File name FULLTEXT01.pdfFile size 913 kBChecksum SHA-512
d1aa4bbb83c58f5cb9db1c05f733649ac086efebcd5d213bc194c2cd26556d3b64befc40dc6409a15c5116c2a4cd12f6e03aea80c229270acf4d33b34fee3ea6
Type fulltextMimetype application/pdf

Other links

Publisher's full textPubMed

Search in DiVA

By author/editor
Fang, Xiaotian T.Sehlin, DagLannfelt, LarsSyvänen, StinaHultqvist, Greta
By organisation
GeriatricsDepartment of Pharmaceutical Biosciences
In the same journal
Biological Procedures Online
Biochemistry and Molecular Biology

Search outside of DiVA

GoogleGoogle Scholar
Total: 8 downloads
The number of downloads is the sum of all downloads of full texts. It may include eg previous versions that are now no longer available

doi
pubmed
urn-nbn

Altmetric score

doi
pubmed
urn-nbn
Total: 103 hits
CiteExportLink to record
Permanent link

Direct link
Cite
Citation style
  • apa
  • ieee
  • modern-language-association-8th-edition
  • vancouver
  • Other style
More styles
Language
  • de-DE
  • en-GB
  • en-US
  • fi-FI
  • nn-NO
  • nn-NB
  • sv-SE
  • Other locale
More languages
Output format
  • html
  • text
  • asciidoc
  • rtf