title: A dual-chamber system for screening cytotoxic effects of hybridoma-produced antibodies.
abstract: There are many methods for evaluating the cytotoxic effect of monoclonal antibodies (MAbs) against cancer cells. Most of these methods require either purified MAbs or biological solutions (e.g., cell culture supernatants, ascitic fluids) containing high concentrations of MAbs. This makes the primary screening of antibody-producing hybridomas for specific cytotoxic antibodies a challenging task. Addressing this issue, this work introduces a high throughput screening method, which enables the identification of cytotoxic antibodies using primary hybridoma populations without prior antibody concentration and/or purification. The method is comprised of a dual-chamber system, where antibody-producing hybridomas and target cancer cells are co-cultured but separated by a porous membrane in which the pore size is sufficient for the diffusion of antibody molecules. The MAbs produced in the system continuously diffuse through the membrane between the two chambers and interact with the target cells placed on the other side of a membrane, resulting in death or proliferation arrest of these cells, if MAbs are cytotoxic or cytostatic. The cytotoxic/cytostatic effect can be registered by measuring the viability of target cells. The advantage of this method is that purification or concentration of antibodies secreted by hybridomas is not required. In addition, this method does not require MAb-secreting hybridomas, which are subcloned or have a high level of MAb production. The method may serve as an effective primary high throughput screening for cytotoxic antibodies.
authors.affiliation: Division of Experimental Therapeutics, Department of Medicine, College of Physicians and Surgeons, Columbia University in New York City, New York 10032, USA.
authors.forename: Ton That Ai
authors.initials: TT
authors.lastname: Long
authors.forename: Gavreel
authors.initials: G
authors.lastname: Kalantarov
authors.forename: Alexandra
authors.initials: A
authors.lastname: Chudner
authors.forename: Ilya
authors.initials: I
authors.lastname: Trakht
mesh_terms: D000911:Antibodies, Monoclonal / Q000032:analysis* / Q000276:immunology / Q000302:isolation & purification; D000951:Antigens, Neoplasm / Q000276:immunology*; D001943:Breast Neoplasms / Q000276:immunology; D003602:Cytotoxicity, Immunologic / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D005434:Flow Cytometry; D057166:High-Throughput Screening Assays; D006801:Humans; D006825:Hybridomas / Q000276:immunology*; D008175:Lung Neoplasms / Q000276:immunology; D008567:Membranes, Artificial; D010051:Ovarian Neoplasms / Q000276:immunology; D014407:Tumor Cells, Cultured
chemical_list: D000911:Antibodies, Monoclonal; D000951:Antigens, Neoplasm; D008567:Membranes, Artificial