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

CE Hagemeyer; K Alt; APR Johnston; GK Such; HT Ta; MKM Leung; S Prabhu; X Wang; F Caruso; K Peter

Journal article

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

CE Hagemeyer, K Alt, APR Johnston, GK Such, HT Ta, MKM Leung, S Prabhu, X Wang, F Caruso, K Peter

Nature Protocols | Published : 2015

DOI: 10.1038/nprot.2014.177

Download PDF

Abstract

Antibody fusion to nonprotein materials such as contrast agents or radio-tracers, nano-or microparticlesor 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 (scF..

View full abstract

University of Melbourne Researchers

Georgina Such Author

Xiaowei Wang Author

Frank Caruso Author

Karlheinz Peter Author

Related Projects (3)

COLORECTAL CANCER - MOLECULAR BASIS TO TARGETED THERAPEUTICS

Cancer of the colon and rectum is the most common form of cancer in Australia. Over 12,000 people are diagnosed each year with colorectal ca..

Engineering and Assembly of Bioinspired Nanostructured Materials

Scientific and technological advances at the frontiers of nano- and bio-technology are poised to revolutionise healthcare and medicine. This..

BUILDING MOLRCULARY ENGINEERED POLYMER NANOMATERIALS

The development of new technologies at the interface between nano- and biotechnology promises to revolutionise healthcare and medicine. This..

Grants

Awarded by National Health and Medical Research Council

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.