Anti-mouse and anti-rabbit IgG secondary VHH single domain antibodies optimized for all immuno-labeling applications
Scientists from the Max-Planck Institute of Biophysical Chemistry in Göttingen have developed anti-IgG secondary VHH single domain antibodies (sdAbs) that outperform commercially available anti-IgG secondary antibodies with regard to sensitivity and specificity and thus offer a sustainable alternative (Figure 1) (ref. 1). A major advantage of the new anti-IgG VHH sdAbs is their easy, cost-effective and animal-friendly production at large scale by expression in E. coli. VHH sdAbs are originally derived from camelid heavy chain antibodies and have a molecular weight of approximately 13 kDa. The VHH sdAbs developed here recognize all mouse IgG subclasses and the sole class of rabbit IgG. Due to their recombinant nature, they can be easily fused to affinity tags or reporter enzymes, such as peroxidases.
Moreover, the VHH sdAbs can be easily linked to all fluorophores via maleimide chemistry and can even be coupled to up to three dyes (ref. 2). The site-specific labeling with multiple fluorophores creates bright imaging reagents suitable for high-resolution confocal and super-resolution microscopy. Since VHH sdAbs are much smaller in size than secondary antibodies they can place fluorophores closer than 2 nm to the primary antibody and thus are perfectly suited for super-resolution imaging. Our scientists have rigorously tested and optimized these VHH sdAbs for all common immuno-labeling approaches. Due to their monovalent nature, these VHH sdAbs do not crosslink primary antibodies and thus allow simpler one-step immuno-staining protocols (pre-incubation of primary antibody with VHH sdAbs). This even enables multi-target localization with primary IgG antibodies from the same species and of the same IgG subclass (Figure 2) (ref. 1).
Taken together, this technology has the potential to replace conventional anti-mouse or anti-rabbit IgG secondary antibodies and it overcomes the lack of signal strength in available alternatives. Currently, we are looking for partners who are interested in in-licensing this technology.
A European priority application has been filed on September 2017.
- Pleiner T, Bates M. and Görlich D., 2017. Available online at bioRxiv: https://doi.org/10.1101/209742
- Pleiner T. et al., 2015. eLife 2015;4:e11349
- Ref.-No.: 0707-5419-LI (846,1 KiB)
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