Local retention of antibodies in vivo with an injectable film embedded with a fluorogen-activating protein

JOURNAL OF CONTROLLED RELEASE

卷:230

Article

Liu, Wen¹  Saunders, Matthew J.^3,4,5,9  Bagia, Christina¹  Freeman, Eric C.⁶  Fan, Yong^5,7  Gawalt, Ellen S.^2,8  Waggoner, Alan S.^3,4,5  Meng, Wilson S.¹ 

[1] Duquesne Univ, Div Pharmaceut Sci, 600 Forbes Ave,413 Mellon Hall, Pittsburgh, PA 15282 USA

[2] Duquesne Univ, Dept Chem & Biochem, 600 Forbes Ave,413 Mellon Hall, Pittsburgh, PA 15282 USA

[3] Mol Biosensor & Imaging Ctr, Pittsburgh, PA 15213 USA

[4] Carnegie Mellon Univ, Pittsburgh, PA 15213 USA

[5] Carnegie Mellon Univ, Dept Biol Sci, 4400 5th Ave, Pittsburgh, PA 15213 USA

[6] Univ Georgia, Coll Engn, Athens, GA 30602 USA

[7] Allegheny Singer Res Inst, Inst Cellular Therapeut, 320 E N Ave, Pittsburgh, PA 15212 USA

[8] McGowan Inst Regenerat Med, Pittsburgh, PA 15213 USA

[9] JLABS, Glycosensors & Diagnost, 3210 Merryfield Row, San Diego, CA 92121 USA

关键词: Bioaffinity film;    Fluorescent imaging;    Antibody formulation;    Single-chain variable fragment;    Mathematical simulation;    Self-assembling peptide;    Fluorogen-activating protein;    EAK16;    Dimerized light chain-5 (dL5);    Controlled release;    Draining lymph nodes;   

DOI  :  10.1016/j.jconrel.2016.03.032

来源: Elsevier

【 摘 要 】

Herein we report an injectable film by which antibodies can be localized in vivo. The system builds upon a bifunctional polypeptide consisting of a fluorogen-activating protein (FAP) and a beta-fibrillizing peptide (beta FP). The FAP domain generates fluorescence that reflects IgG binding sites conferred by Protein A/G (pAG) conjugated with the fluorogen malachite green (MG). A film is generated by mixing these proteins with molar excess of EAK16-II, a beta FP that forms beta-sheet fibrils at high salt concentrations. The IgG-binding, fluorogenic film can be injected in vivo through conventional needled syringes. Confocalmicroscopic images and dose-response titration experiments showed that loading of IgG into the film was mediated by pAG(MG) bound to the FAP. Release of IgG in vitro was significantly delayed by the bioaffinity mechanism; 26% of the IgG were released from films embedded with pAGMG after five days, compared to close to 90% in films without pAGMG. Computational simulations indicated that the release rate of IgG is governed by positive cooperativity due to pAGMG. When injected into the subcutaneous space of mouse footpads, film-embedded IgG were retained locally, with distribution through the lymphatics impeded. The ability to track IgG binding sites and distribution simultaneously will aid the optimization of local antibody delivery systems. (C) 2016 Elsevier B.V. All rights reserved.

【 授权许可】

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