Journal article

Particle generation, functionalization and sortase A-mediated modification with targeting of single-chain antibodies for diagnostic and therapeutic use

Christoph E Hagemeyer, Karen Alt, Angus PR Johnston, Georgina K Such, Hang T Ta, Melissa KM Leung, Sandeep Prabhu, Xiaowei Wang, Frank Caruso, Karlheinz Peter

NATURE PROTOCOLS | NATURE PUBLISHING GROUP | Published : 2015

Abstract

Antibody fusion to nonprotein materials such as contrast agents or radio-tracers, nano- or microparticles or small-molecule drugs is attracting major interest for molecular imaging and drug delivery. Nondirected bioconjugation techniques may impair antibody affinity, result in lower amounts of functional antibodies and generate multicomponent mixtures. We present a detailed protocol for the enzymatic bioconjugation of small recombinant antibodies to imaging particles, and we also describe the generation of and conjugation to a low-fouling capsule assembled for drug delivery from PEG and PVPON (poly(N-vinylpyrrolidone) by a layer-by-layer (LbL) technique. The single-chain variable fragment (s..

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Grants

Awarded by National Health and Medical Research Council


Awarded by National Heart Foundation


Awarded by Australian Research Council (ARC) Discovery Project


Awarded by National Heart Foundation Career Development Fellowship


Awarded by German Research Foundation


Awarded by ARC Future Fellowship


Awarded by National Heart Foundation Postdoctoral Fellowship


Awarded by ARC Federation Fellowship


Funding Acknowledgements

This work was funded by the National Health and Medical Research Council (grant nos. 472665, 472667 and 487922), the National Heart Foundation (grant no. G09M4345) and an Australian Research Council (ARC) Discovery Project (DP0877360). The work was supported in part by the Victorian Government's Operational Infrastructure Support Program. C.E.H. is supported by a National Heart Foundation Career Development Fellowship (CR 11M 6066), K.A. is supported by the German Research Foundation (Al 1521/1-1), A.P.R.J. is supported by an ARC Future Fellowship (FT110100265), H.T.T. is supported by a National Heart Foundation Postdoctoral Fellowship (PF09M4688), M.K.M.L. is supported by an ARC Australian Postgraduate Award, F.C. is supported by an ARC Federation Fellowship (FF0776078) and K.P. is supported by an ARC Future Fellowship (FT0992210). We thank G. Krippner for his help with Figure 1.