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Elife. 2017 Jun 27;6. pii: e24903. doi: 10.7554/eLife.24903.

Affimer proteins are versatile and renewable affinity reagents.

Author information

1
School of Molecular and Cellular Biology, University of Leeds, Leeds, United Kingdom.
2
Astbury Centre for Structural and Molecular Biology, University of Leeds, Leeds, United Kingdom.
3
Leeds Institute of Cancer Studies and Pathology, University of Leeds, Leeds, United Kingdom.
4
The Pirbright Institute, Woking, United Kingdom.
5
Central Laser Facility, Research Complex at Harwell, STFC Rutherford Appleton Laboratory, Didcot, United Kingdom.
6
Avacta Life Sciences, Wetherby, United Kingdom.
7
Leeds Institute of Biomedical and Clinical Sciences, University of Leeds, Leeds, United Kingdom.
8
Leeds Institute of Cardiovascular and Metabolic Medicine, University of Leeds, Leeds, United Kingdom.
9
DSTL Porton Down, Salisbury, United Kingdom.
10
Oxford Protein Production Facility UK, Research Complex at Harwell, STFC Rutherford Appleton Laboratory, Didcot, United Kingdom.
11
School of Chemistry, University of Leeds, Leeds, United Kingdom.
12
School of Biomedical Sciences, University of Leeds, Leeds, United Kingdom.

Abstract

Molecular recognition reagents are key tools for understanding biological processes and are used universally by scientists to study protein expression, localisation and interactions. Antibodies remain the most widely used of such reagents and many show excellent performance, although some are poorly characterised or have stability or batch variability issues, supporting the use of alternative binding proteins as complementary reagents for many applications. Here we report on the use of Affimer proteins as research reagents. We selected 12 diverse molecular targets for Affimer selection to exemplify their use in common molecular and cellular applications including the (a) selection against various target molecules; (b) modulation of protein function in vitro and in vivo; (c) labelling of tumour antigens in mouse models; and (d) use in affinity fluorescence and super-resolution microscopy. This work shows that Affimer proteins, as is the case for other alternative binding scaffolds, represent complementary affinity reagents to antibodies for various molecular and cell biology applications.

KEYWORDS:

Affimer; E. coli; antibody; assay; biochemistry; cell biology; human; mouse

PMID:
28654419
PMCID:
PMC5487212
DOI:
10.7554/eLife.24903
[Indexed for MEDLINE]
Free PMC Article

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