Quantitative spatiotemporal analysis of antibody fragment diffusion and endocytic consumption in tumor spheroids.
Antibody-based cancer treatment depends upon distribution of the targeting macromolecule throughout tumor tissue, and spatial heterogeneity could significantly limit efficacy in many cases. Antibody distribution in tumor tissue is a function of drug dosage, antigen concentration, binding affinity, antigen internalization, drug extravasation from blood vessels, diffusion in the tumor extracellular matrix, and systemic clearance rates. We have isolated the effects of a subset of these variables by live-cell microscopic imaging of single-chain antibody fragments against carcinoembryonic antigen in LS174T tumor spheroids. The measured rates of scFv penetration and retention were compared with theoretical predictions based on simple scaling criteria. The theory predicts that antibody dose must be large enough to drive a sufficient diffusive flux of antibody to overcome cellular internalization, and exposure time must be long enough to allow penetration to the spheroid center. The experimental results in spheroids are quantitatively consistent with these predictions. Therefore, simple scaling criteria can be applied to accurately predict antibody and antibody fragment penetration distance in tumor tissue.
Thurber
Greg M
GM
Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Wittrup
K Dane
KD
D000906:Antibodies / Q000378:metabolism; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000473:pathology / Q000628:therapy; D004058:Diffusion; D004705:Endocytosis / Q000502:physiology*; D049452:Green Fluorescent Proteins / Q000378:metabolism / Q000493:pharmacokinetics; D006801:Humans; D007128:Immunoglobulin Fragments / Q000378:metabolism; D007167:Immunotherapy; D008954:Models, Biological; D011993:Recombinant Fusion Proteins / Q000378:metabolism / Q000493:pharmacokinetics; D018874:Spheroids, Cellular / Q000378:metabolism*; D013997:Time Factors; D014018:Tissue Distribution; D014407:Tumor Cells, Cultured
D000906:Antibodies; D007128:Immunoglobulin Fragments; D011993:Recombinant Fusion Proteins; D049452:Green Fluorescent Proteins