A method for phospholipid-mediated delivery of specific antibodies into adherent cultured cells.
A simple and rapid procedure for the intracellular delivery of macromolecules into adherent cultured cells is described. Cells are incubated with cold glycerol, then transiently made permeable with L-alpha-lysophosphatidylcholine (LPC) in the presence of test compound to be loaded into cells. LPC induces temporary permeability of the plasma membrane, as evidenced by the loss and recovery of the cells' ability to exclude trypan blue. Molecules at least as large as antibodies are internalized during this transient permeability. Antibodies delivered intracellularly in this manner are able to complex with their specific antigen and exert functional consequences on normal cell metabolism, suggesting that this procedure is useful for determining protein function. As one example of this, we present data on the ability of specific antibodies, delivered intracellularly in this manner, to inhibit morphological differentiation (i.e., neurite outgrowth) in a neuroblastoma cell line.
Shea
T B
TB
Ralph Lowell Laboratories, Mailman Research Center, McLean Hospital, Belmont, MA 02178.
Beermann
M L
ML
D000906:Antibodies / Q000008:administration & dosage* / Q000378:metabolism / Q000494:pharmacology; D001369:Axons / Q000502:physiology / Q000648:ultrastructure; D003994:Bucladesine / Q000494:pharmacology; D002448:Cell Adhesion; D002454:Cell Differentiation / Q000187:drug effects; D002463:Cell Membrane Permeability / Q000187:drug effects*; D002470:Cell Survival; D008244:Lysophosphatidylcholines / Q000494:pharmacology*; D008869:Microtubule-Associated Proteins / Q000276:immunology; D009447:Neuroblastoma; D014404:Tubulin / Q000276:immunology / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D000906:Antibodies; D008244:Lysophosphatidylcholines; D008869:Microtubule-Associated Proteins; D014404:Tubulin; D003994:Bucladesine