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“Optimizing Blood-Brain Barrier Transcytosis: Shorter Linker Enhances TfR-Mediated Transport in Bispecific Antibody RmAb158-scFv8D3”

# Optimizing Blood-Brain Barrier Transcytosis: Shorter Linker Enhances TfR-Mediated Transport in Bispecific Antibody RmAb158-scFv8D3

The blood-brain barrier (BBB) is a highly selective and tightly regulated interface that protects the brain from harmful substances while allowing the transport of essential nutrients. However, this protective mechanism also poses a significant challenge for delivering therapeutic agents to the brain, particularly in the treatment of neurodegenerative diseases such as Alzheimer’s disease (AD). Recent advances in antibody engineering have focused on optimizing the transcytosis of therapeutic molecules across the BBB via receptor-mediated transport (RMT). One promising approach involves the use of bispecific antibodies that target both a therapeutic antigen and a BBB transport receptor, such as the transferrin receptor (TfR). A recent study highlights the role of linker length in enhancing the efficiency of TfR-mediated transport, specifically in the bispecific antibody RmAb158-scFv8D3.

## The Challenge of BBB Drug Delivery

The BBB is formed by endothelial cells connected by tight junctions, supported by pericytes and astrocytic end-feet. This structure restricts the passage of large molecules, including most therapeutic antibodies, into the brain. While small lipophilic molecules can diffuse across the BBB, larger biologics require active transport mechanisms. RMT, which leverages endogenous receptors such as TfR, insulin receptor, and low-density lipoprotein receptor-related protein 1 (LRP1), has emerged as a promising strategy for delivering biologics to the brain.

TfR is particularly attractive for RMT because it is highly expressed on BBB endothelial cells and facilitates the transport of iron-bound transferrin into the brain. Bispecific antibodies that bind to TfR and a therapeutic target in the brain can exploit this pathway to cross the BBB. However, the design of such antibodies must balance high affinity for TfR (to ensure efficient transport) with the need to avoid receptor saturation and lysosomal degradation.

## RmAb158-scFv8D3: A Bispecific Antibody for Alzheimer’s Disease

RmAb158-scFv8D3 is a bispecific antibody designed to target amyloid-beta (Aβ) plaques, a hallmark of Alzheimer’s disease, while simultaneously engaging TfR for BBB transcytosis. The antibody consists of two functional domains: RmAb158, which binds to Aβ, and scFv8D3, a single-chain variable fragment that binds to TfR. This dual-targeting approach allows the antibody to cross the BBB and accumulate in the brain, where it can neutralize Aβ aggregates.

Despite its promise, the efficiency of RmAb158-scFv8D3 transport across the BBB is influenced by several factors, including the affinity of scFv8D3 for TfR and the structural design of the antibody. One critical design parameter is the length of the linker connecting