In vitro methods like our In Vitro Antibody Pharmacokinetics Assay offer more accurate results than traditional animal studies. The assay utilizes tumor spheroid models to measure the distribution of biologicals into solid tumor models in vitro. The protocol involves generating free-floating tumor spheroids, treating them with antibody therapeutics at various concentrations, fixing them at specified time points, labeling the antibody with fluorescent secondary to label therapeutic antibody, applying tissue clearing to render tumor spheroids transparent, high content confocal imaging, and analysis of resultant images to assess distance and quantity of antibody penetration into tumor spheroid.
The test article concentration involves an 8-point assay and a single point assay, with custom concentrations available. Time points include 0.5 hr, 1 hr, 2 hr, 4 hr, 8 hr, 24 hr, 48 hr, with custom time points available. Three replicates per concentration are required, with 0.5% DMSO as the vehicle control and anti-β-integrin as the positive control. Antibody concentration is measured as a mean of pixel intensity in concentric bands of decreasing radius from the edge of the tumor spheroid into the center.
The resultant data includes distribution curves, velocity curves, TD50 (time to distribute to 50% of total) values, and concentration-time curves (measured as AUC). Representative data includes confocal sections from the center of HepG2 tumor spheroids depicting the relative distribution of anti-β-integrin antibody at various concentrations and times, a representative curve depicting parameters used for quantitation of antibody distribution into tumor spheroids, and quantitative measurements of the extent of distribution of antibody into tumor spheroids. Antibody distribution curve depicts penetration of antibody into tumor spheroid at varying doses over time. Dose and time dependence of velocity of antibody distribution and time to distribute to 50% of spheroid radius (DT50) and maximum distance of penetration are also measured.
The Advantages
There are several advantages over traditional animal studies.
- It is more efficient and cost-effective, as it can test multiple compounds simultaneously and does not require the use of animals.
- It provides more accurate results, as it utilizes tumor spheroid models that better reflect human physiology.
- It can be customized to meet the specific needs of each client, allowing for greater flexibility in drug development.
Get Started Today
The future of antibody pharmacokinetics lies in in vitro methods like our In Vitro Antibody Pharmacokinetics Assay. These methods offer a more efficient and accurate alternative to traditional animal studies and are becoming increasingly popular in the pharmaceutical industry. Visikol’s expertise in the field of in vitro methods for drug development makes them an ideal partner for pharmaceutical companies looking to accelerate the drug discovery and development process. To learn more about Visikol’s In Vitro Antibody Pharmacokinetics Assay, reach out to a member of our team.