Affimer proteins are versatile and renewable affinity reagents
Tiede C., Bedford R., Heseltine SJ., Smith G., Wijetunga I., Ross R., AlQallaf D., Roberts APE., Balls A., Curd A., Hughes RE., Martin H., Needham SR., Zanetti-Domingues LC., Sadigh Y., Peacock TP., Tang AA., Gibson N., Kyle H., Platt GW., Ingram N., Taylor T., Coletta LP., Manfield I., Knowles M., Bell S., Esteves F., Maqbool A., Prasad RK., Drinkhill M., Bon RS., Patel V., Goodchild SA., Martin-Fernandez M., Owens RJ., Nettleship JE., Webb ME., Harrison M., Lippiat JD., Ponnambalam S., Peckham M., Smith A., Ferrigno PK., Johnson M., McPherson MJ., Tomlinson DC.
<jats:p>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.</jats:p>