Method for isolating cells

Claim 3

A method according to claim 1 wherein the complex formed with the first substance establishes a barrier condition within the vicinity of the desired cells and said second substance responds to the barrier condition and interacts with all desired cells in a manner which is different from its interaction with all undesired cells, the interaction of the second substance rendering possible the selection of the desired cells.

Claim 4

A method according to claim 1 wherein the second substance has affixed thereto a third substance which is drug capable of destroying a living cell upon entry therein, and said-complex establishes a barrier condition within the immediate vicinity of the desired cells which prevents the second substance from contacting and destroying the desired cells thereby selecting the desired cells and destroying the undesired cells.

Claim 5

A method according to claim 1 wherein the first substance is scavenger cells which have been subjected to a treatment causing their death after a given amount of time of normal metabolism,and which have a site capable of specifically binding with said product to form said complex and which have an enhanced ability to take up molecules from their environment when the cells are bound as a complex; said scavenger cells are combined with said cell population before its placement in the predetermined environment; the predetermined environment separates the position of each cell of the population and the scavenger cells with respect to the other cells of the population and the scavenger cells, the scavenger cells essentially surround each cell in the population, and said cellular product from each desired cell from which it is secreted is inhibited from migrating such that only the scavenger cells in the immediate vicinity of a desired cell encounter said cellular product: sufficient time for said cellular product to be secreted by the desired cells and to bind with said scavenger cells within the immediate vicinity of the desired cell is allowed; and - the second substance is a material normally lethal to said cell population and scavenger cells, the complex between said product and said scavenger cells causes the scavenger cells in the immediate vicinity of a desired cell to take up said second substance to a degree sufficient to prevent contact of the desired cells and the second substance, thereby preventing death of the desired cells and selecting the desired cells; the undesired cells are killed by the lethal environment; and the scavenger cells eventually die as a result of their pretreatment.

Claim 6

A method according to claim 1 wherein the first substance is carried on carrier particles and said particles are combined with the cell population before it is placed in the predetermined environment; the carrier particles essentially surround the cells of the population when combined with the predetermined environment: the second substance is affixed to an enzyme, is capable of specifically binding with free first substance bound in said complex, and preferentially directs the enzyme to the vicinity of undesired cells; or the second substance is affixed to an enzyme binds only with said cellular product or said complex of carrier particle and cellular product; any unbound second substance is removed from the population of mixed cells; and if the second substance binds with free first substance, a third substance is added to the population of cells containing said second substance, which third substance is capable of being converted by said enzyme to a lethal, colored, colorless, fluorescent or insoluble compound which selectively acts upon the undesired cells and produces selection of the desired cells; or, if the second substance binds with said product or said complex, a lethal, third substance is added to the population of cells containing said second substance which third substance is converted by said enzyme to a nonlethal compound and the desired cells are selected by rescue from said lethal third substance while the undesired cells are killed.

Claim 7

A method according to claim 1 wherein the first substance is carried on carrier particles, said particles are combined with the cell population before it is placed in the predetermined environment and said particles are selected from red blood cells and liposomes which contain an enzyme: the carrier particles essentially surround the cells of the population when it is placed in the predetermined environment: only the carrier particles in the immediate vicinity of the desired cells encounter and complex with the cellular product; the second substance specifically binds only to the complex of particles and product, and causes the particles of said complexes to lyse and release said enzyme: a lethal, soluble or heavy third substance is added to the cell population containing released enzyme. ¡said third substance is converted by the enzyme to a nonlethal compound if it is lethal, an insoluble precipitate if it is soluble or a light compound if it is heavy, and the desired cells are selected by rescue from a lethal environment if the third substance is lethal, by precipitation if the third substance is soluble. or flotation if the third substance is heavy.

Claim 8

A method according to claim 1 wherein the first substance is carried on carrier particles, said particles are combined with the cell population before it is placed in the predetermimed environment and said particles are selected from red blood cells and liposomes which contain a rescue chemical; the carrier particles essentially surround the cells of the population when combined with the predetermined environment- only the carrier particles within the immediate vicinity of the desired cells encounter and complex with the cellular product: the second substance specifically binds only to the complex of particles and product, and causes the particles of said complexes to lyse and release said rescue chemical only within the vicinity of the desired cells causing a barrier around the desired cells; a lethal third substance is added to the cell population containing released rescue chemical, the desired cells are rescued from the effects of the third substance by the barrier of rescue chemical and the undesired cells are killed by the third substance.

Claim 9

A method according to claim 1 wherein the predetermined environment is hollow, semipermeable beads sized for retention of the cells of the population and permitting the diffusion of the first and second substances through the bead walls: the second substance carries an enzyme and will bind only to said complex or will bind only to uncomplexed first substance; unbound second substance is removed from the beaded population of cells; and the second substance bound to the complex interacts with the bead material of the beads containing the desired cells, or the second substance bound to the first substance alone interacts with the bead material of the beads containing the undesired cells; a third bead material liquifying substance is added to cause a distinguishable condition selected from liquification, or maintanence of the gel nature of the bead material thereby producing selection of the desired cells.

Claim 10

A method according to claim 1 wherein the first substance is carried on carrier particles, said particles are combined with the cell population before it is placed in the predetermined environment, and said particles are selected from red blood cells and liposomes and essentially surround the cells of the population when combined in the predetermined environment: the predetermined environment is hollow, semipermeable beads sized for retention of the cells of the population; only the carrier particles within the beads containing the desired cells encounter and complex with the cellular product; the bead wall material is permeable to the second substance; the second substance binds only to the complex of carrier particles and cellular product and causes lysis of the carrier particles only when it is bound to the complex or the second substance is an enzyme bound to complex compliment: unbound second substance is removed from the beads: when the second substance causes lysing action, lysate is released from the carrier particles which escapes through the semipermeable walls of the beads and causes the beads containing the desired cells to rise above the level of the other beads; or when the second substance is an enzyme bound to complement, a third substance is added to the cell population which is converted by said enzyme to an agent which lyses the carrier particles and releases lystate therefrom which escapes through the semipermeable walls of the beads and causes the beads containing the desired cells to rise above the level of the other beads.

Claim 11

A method according to claim 1 wherein the cellular product is a first antibody, the first substance is a combination of an antigen specific for said antibody and the Fab fragments of a second antibody exhibiting affinity for said antigen, the first substance and the cell population are encapsulated within a semi-permeable encapsulating material which is the predetermined environment, sufficient time is allowed for the first antibody secreted by the desired cells within the encapsulated material to displace any of the Fab fragments exhibiting less affinity for said antigen than exhibited by said first antibody; the second substance is a material which binds to the Fe region of said displacing first antibody and which directly or indirectly causes a condition within the capsules containing said first antibody which distinguishes such capsules from all other capsules.