July 2018
Volume 59, Issue 9
Open Access
ARVO Annual Meeting Abstract  |   July 2018
Hyaluronic acid conjugation significantly extends the vitreal half-life of antibody fragments
Author Affiliations & Notes
  • Susan Crowell
    Preclinical and Translational PKPD, Genentech, South San Francisco, California, United States
  • Amin Famili
    Drug Delivery, Genentech, South San Francisco, California, United States
  • C. Andrew Boswell
    Preclinical and Translational PKPD, Genentech, South San Francisco, California, United States
  • Joyce Chan
    Biochemical and Cellular Pharmacology, Genentech, South San Francisco, California, United States
  • Julia Gray
    Biochemical and Cellular Pharmacology, Genentech, South San Francisco, California, United States
  • Kelly Loyet
    Biochemical and Cellular Pharmacology, Genentech, South San Francisco, California, United States
  • Laetitia Comps-Agrar
    Biochemical and Cellular Pharmacology, Genentech, South San Francisco, California, United States
  • Amrita kamath
    Preclinical and Translational PKPD, Genentech, South San Francisco, California, United States
  • Karthikan Rajagopal
    Drug Delivery, Genentech, South San Francisco, California, United States
  • Footnotes
    Commercial Relationships   Susan Crowell, Genentech (E); Amin Famili, Genentech (E); C. Andrew Boswell, Genentech (E); Joyce Chan, Genentech (E); Julia Gray, Genentech (E); Kelly Loyet, Genentech (E); Laetitia Comps-Agrar, Genentech (E); Amrita kamath, Genentech (E); Karthikan Rajagopal, Genentech (E)
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science July 2018, Vol.59, 235. doi:
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      Susan Crowell, Amin Famili, C. Andrew Boswell, Joyce Chan, Julia Gray, Kelly Loyet, Laetitia Comps-Agrar, Amrita kamath, Karthikan Rajagopal; Hyaluronic acid conjugation significantly extends the vitreal half-life of antibody fragments. Invest. Ophthalmol. Vis. Sci. 2018;59(9):235.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose : Treatment of ocular diseases associated with neovascularization requires frequent intravitreal injection of anti-vascular endothelial growth factor (aVEGF) therapies. Reducing the required frequency of aVEGF injections—and associated clinical visits—may improve patient adherence to the prescribed treatment regimen and improve outcomes. Frequent re-dosing of biologic aVEGF therapies is dictated by their clearance from the vitreous humor; strategies to modify vitreal clearance rates may extend dosing intervals. In this work, we explore conjugation of fragment antibodies (Fab) to the biopolymer hyaluronic acid (HA) as a half-life modifying strategy and assess the impact on Fab biophysical properties and vitreal pharmacokinetics.

Methods : Two model Fab molecules—one species-matched rabbit Fab (rabFab) used for rabbit PK experiments and one human aVEGF Fab (Fab1) used for biophysical characterization—were expressed with C-terminal cysteine residues and conjugated to maleimide-modified linear HA of varying molecular weights. HA-Fab1 conjugates were characterized for solution physical properties, melting behavior, and higher-order structure. HA-rabFab conjugates of three distinct molecular weights and hydrodynamic radii (RH) were assessed for in vivo pharmacokinetic performance relative to unconjugated rabFab after intravitreal injection in rabbits.

Results : Covalent conjugation of Fab1 to HA did not measurably alter the biophysical properties of the protein: melting temperature (Fab1: 90.9°C; HA-Fab1: 90.4°C), and secondary and tertiary structure measured by circular dichroism were unchanged. Conjugation to HA significantly slowed the in vivo clearance of rabFab from the rabbit vitreous in a RH-dependent manner. Compared to free rabFab (observed vitreal half-life of 2.8 days), HA-rabFab conjugates cleared with observed half-lives of 7.6, 10.2 and 18.3 days for 40 kDa, 200 kDa and 600 kDa HA conjugates, respectively.

Conclusions : Conjugation of therapeutic Fab molecules to HA resulted in significant half-life extension while maintaining required Fab biophysical properties. This strategy may enable long-acting delivery of proteins targeting posterior ocular diseases.

This is an abstract that was submitted for the 2018 ARVO Annual Meeting, held in Honolulu, Hawaii, April 29 - May 3, 2018.

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