Immunogenicity Testing of Therapeutic Antibodies in Ocular Fluids after Intravitreal Injection

Abstract

The IC assay allows a reliable ADA detection in matrices with high drug concentrations, such as ocular fluids.

Biotherapeutics and immunogenicity: ophthalmic perspective

“…Demonstration of ADA in patient's serum who took intra-vitreal anti-VEGF injections was attributed to be in response to the molecules which entered systemic circulation via aqueous drainage pathways. Intra-vitreal presence of these ADA have now been established and confirms presence of adaptive immunity in the eye in certain pathologies like AMD [14].…”

Use of Ella ® to Facilitate drug Quantification and Antidrug Antibody Detection in Preclinical Studies

“…In contrast, an increased dynamic range would be beneficial because samples would not require high dilutions to be in the assay range. In addition, lower sample consumption of the Ella platform (2.5 vs 25 μl in ELISA) would reduce the difference in sensitivity for samples with limited availability such as mouse samples or rare sample matrices like eye, synovial or cerebrospinal fluids [1][2][3][4]. For example, if only 2.5 μl is available, the sensitivity of the Ella assay would be still 15 ng/ml, whereas the sensitivity of ELISA would be decreased to 19 ng/ml.…”

Selected Aspects of Ocular Toxicity Studies With a Focus on High-Quality Pathology Reports: A Pathology/Toxicology Consultant’s Perspective

“…Hurdles with poor drug tolerance and related false-negative results with state-of-the-art bridging assays to detect ADA in ocular fluid samples have recently been overcome with more sensitive immune complex assays. 47 Immune complex assays of VH and AH samples with high residual drug concentrations following intravitreal injection in minipigs and monkeys detected ADA, although systemic ADA were detected earlier than in ocular fluids and systemic ADA response was not always accompanied by the presence of ADA in ocular fluids, indicating systemic ADA detection is adequate in most cases for ocular toxicity studies. Aqueous humor was a good predictive matrix for ADA detection in VH, which is important for ease of sampling in AH compared to VH in ocular toxicity studies and minimizing potential contamination issues in VH.…”

“…Recent publications with relevance to ocular toxicity studies in minipigs, primarily Göttingen minipigs, include basic ocular histomorphometry, 41 pre- and postnatal development of the eye, 42 ocular immunohistochemistry and in situ hybridization biomarkers, 43 electroretinography (ERG), 44 and optical coherence tomography (OCT). 43,45 Immunology 46 and ocular immunogenicity testing 47 of minipigs has been characterized, and although the minipig may be suitable for biologics, this species can mount a substantial immunogenic response to human proteins. 13 Vitreous volume in the Gottingen minipig eye using a frozen eye collection technique by Struble et al 48 was approximately 2.3 mL, which is important for calculating a human equivalent dose (HED) and corresponding ocular dose safety margins for intravitreal drugs in this species.…”