Human-Based Antibody Therapeutic Detection

Brain-wide assay for measuring the target engagement, efficacy, and distribution of novel antibody therapeutics.

Background

The blood-brain barrier (BBB) is a selective semipermeable membrane that separates the circulating blood from the brain parenchyma where the neurons and glial cells that are the targets of therapeutics reside. One of the major challenges in developing effective treatments for neurological disorders is engineering therapeutic agents to cross the BBB and reach their targets within the CNS. This scientific challenge has led to a new focus on the creation of human-based antibody therapeutics.

With the recent approval of two antibody therapeutics for Alzheimer's Disease, a large effort is underway to generate next-generation antibodies that harness 'brain shuttle' mechanisms to cross the BBB in the brain parenchyma. Consequently, even with many promising vectors, many potential antibody-based therapeutics are still unable to cross the blood brain barrier efficiently. Consequently, we have developed our Human-Based Antibody Therapeutic Detection Full-pipeline service. This all encompassing service is well suited for researchers investigating the distribution, efficacy, and target engagement of their novel human-based antibody therapeutics.

Learn more about our work with Human-Based Antibody Therapeutics and the Blood Brain Barrier on our science page →

Human-Based Antibody Therapeutic Detection

Full-Pipeline

From whole brains to actionable readouts of therapeutic distribution, efficacy, and target engagement

Experimental Treatments

Perform your favorite behavioral, pharmacological, genetic, or other manipulation, and send us your samples. Or collaborate with us, and we’ll perform the animal treatments for you.

Whole-Brain Tissue Clearing

The Human-Based Antibody Therapeutic Detection Full-Pipeline service employs Translucence Biosystems’ modified iDISCO+ tissue clearing protocol to render intact brain samples optically transparent. In combination with an Anti-Human secondary antibody, Translucence Biosystems generates snapshots of the distribution of your therapeutics across the whole brain.

Light Sheet Imaging

Cleared, labeled samples are imaged on the Lightsheet Z.1 equipped with the Mesoscale Imaging System™. The Mesoscale Imaging System is optimized for imaging large intact tissues with unprecedented speed and resolution and is compatible with the high refractive index solutions used in tissue clearing protocols, such as iDISCO+.

Translucence’s imaging pipelines capture cellular-resolution, whole-brain images providing actionable readouts of therapeutic distribution, efficacy, and target engagement.

Light Sheet Image Stitching

We employ our in-house developed software solution, Stitchy™, for automated light sheet image stitching. Stitchy is a robust software solution that stitches large imaging datasets 2.5-5.5 times faster than leading commercial software solutions. Stitchy enables seamless integration with downstream image analysis pipelines by eliminating intermediate file formats.

AI-Powered Quantification

The Translucence Teravoxel Tool Kit (3TK) software applies deterministic filters and AI-powered segmentation to provide actionable readouts of brain-wide therapeutic biodistribution, efficacy, and target engagement.

Actionable data

Browse actionable insights generated by the

Human-Based Antibody Therapeutic Detection

service

Intravenous Administration of Monospecifc and Bispecific anti BACE1 Antibodies

We dosed mice intravenously with various concentrations of a monospecific antibody targeting BACE1 or a bispecific antibody that is similar, but with the addition of a domain binding to the transferrin receptor (TfR1). The transferrin receptor is found in blood vessel endothelial cells where it acts as a carrier to transport iron. Antibodies binding to the transferrin receptor can hijack this mechanism to be transported across the blood-brain barrier.

Staining Validation with Multiple anti Human IgG Antibodies

When developing the methodology for whole-brain imaging of IV-dosed experimental therapeutic human antibodies, we tested multiple different secondary antibodies. The top panels show sagittal planes of intact brains stained with three different secondary antibodies. We determined that the Fcy antibody had better brain penetration and more even whole-brain staining. Importantly, all three secondary antibodies produced qualitatively similar patterns of immunoreactivity.

Enrichment of Bispecific Antibody Throughout the Brain

Our modified iDISCO+ protocol allows for imaging of an entire intact tissue, giving an unbiased snapshot of immunoreactivity throughout the brain. Consistent with the data from the hippocampus, we discovered that the monospecific antibody has negligible brain penetration, whereas the bispecific antibody is identified in both the vasculature and parenchyma throughout the brain. The bispecific BACE1 antibody is particularly enriched in a number of brain areas including the Pontine Gray, Medulla and Glomeruli in the Olfactory Bulb.

iDISCO Whole Brain Imaging of IV dosed anti BACE1 Antibodies

24 hours after dosing, using our modified iDISCO+ protocol, we cleared and stained perfusion-fixed brains using an anti-human IgG secondary antibody. We could not detect any immunoreactivity in the brains of mice injected with the monospecific antibody, with background signals identical to brains from non-infected mice. With the bispecific antibody, strong staining was observed in blood vessels, and importantly, in the brain parenchyma. In independent experiments, we saw similar patterns of staining in the hippocampus of IV-dosed mice stained with traditional slice IHC.

Validation of BACE1 Targeting of the IV-Dosed Antibodies

Traditional slice IHC experiments validated that there is enrichment of the IV-dosed bispecific antibody (50 and 125 mmol/kg) vs. the monospecific antibody (125 mmol/kg) in the dentate gyrus (DG) of the hippocampus (green panels and bar graph). This pattern is very similar to that seen in slices from non-dosed mice stained directly with an anti-BACE1 antibody (red staining).

The Human-Based Antibody Therapeutic Detection Service

The Human-Based Antibody Therapeutic Detection service leverages next-generation 3D histology technologies including tissue clearing and light sheet imaging to provide actionable readouts of brain-wide therapeutic biodistribution, efficacy, and target engagement.

This service is suited for researchers investigating the distribution, efficacy, and target engagement of their novel human-based antibody therapeutics.

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