US 5,646,001 AGranted

Affinity-binding separation and release of one or more selected subset of biological entities from a mixed population thereof

AssigneeImmunivest Corporation

InventorsLeon W. M. M. Terstappen, Galla C. Rao, Dhanesh I. Gohel, Brian P. Feeley, Steven Gross, Ellen S. Church, Paul A. Liberti

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Abstract

A method for separation of a mixture of biological entities into at least three distinct, subpopulations. Different antibodies are provided, with each antibody bound to a solid support in a unique manner such that by a manipulation of the physical or chemical environment, the bonds between the antibodies and the solid supports can be selectively broken. The mixed population of cells is incubated with the antibodies. The cells are magnetically separated from a test medium and collected in a monolayer upon a collection surface. Then by manipulation of the physicochemical environment, specific linkages can be broken and desired cell subpopulations released from the collection surface. This method has medically significant diagnostic and therapeutic applications, as entire cell types can be separated from non-malignant medically vital cell types. Cancer can be diagnosed, staged, and monitored. Genetic analysis from maternal blood, CVS, or amniocentesis samples is possible. Diseases such as AIDS, tuberculosis or hepatitis can be monitored. This invention also has utility in the fields of bone marrow transplantation, fetal cell research, in vitro fertilization, and gene therapy.

Claims38

Dependent claims

A method for isolating a selected subset of biological entities from among a plurality of subsets in a mixed population of said biological entities, said biological entities being selected from the group consisting of cells, bacteria, viruses, or components thereof and toxins, said plurality of subsets comprising a first subset having a first characteristic determinant and at least one other subset having at least a second characteristic determinant, one of said first subset and said at least one other subset being said selected subset, said method comprising the steps of:

(a) providing a plurality of capture agents, each capture agent comprising a receptor attached to a solid support, said plurality of capture agents including a first capture agent comprising a receptor which specifically binds, either directly or indirectly, to said first characteristic determinant and at least one other capture agent comprising a receptor which specifically binds, either directly or indirectly, to said second characteristic determinant, the subset to which each said capture agent binds being its target subset;
(b) contacting a test sample comprising said mixed population of biological entities with said plurality of capture agents, whereby the receptor of said first capture agent specifically binds to said first subset and the receptor of said at least one other capture agent specifically binds to said at least one other subset, at least one of said first and said second capture agents being dissociably linked to its target subset;
(c) collecting on a collection surface the bound subsets from said test sample and separating from said bound subsets any subset of said population than is not bound to a capture agent;
(d) selectively releasing one dissociably linked target subset, among said bound subsets, from said collection surface by dissociating said one dissociably linked subset from all or part of the capture agent to which it is bound;
and
(e) isolating said selectively released target subset.

3 more elements

A method for isolating a selected subset of cells from among a plurality of cell subsets in mixed cell population in a blood sample, said plurality of subsets comprising a first subset having a first characteristic antigen and at least one other subset having at least a second characteristic antigen, one of said first subset and said at least one other subset being said selected subset, said method comprising the steps of:

(a) providing a plurality of capture agents, each capture agent comprising an antibody attached to a solid support, said plurality of capture agents including a first capture agent comprising an antibody which specifically binds, either directly or indirectly, to said first characteristic antigen and at least a one other capture agent comprising an antibody which specifically binds, either directly or indirectly, to said second characteristic antigen, the antibody of each of said first and at least one other capture agent being dissociably attached to its solid support and the attachment of antibody to solid support in said first capture agent being different from the attachment of antibody to solid support in said at least one other capture agent, the subset to which each said capture agent binds being its target subset;
(b) contacting said blood sample with said plurality of capture agents, whereby the antibody of said first capture agent specifically binds to said first subset and the antibody of said at least one other capture agent specifically binds to said at least one other subset;
(c) collecting on a collection surface the bound subsets from said test sample and separating from said bound subsets any subset of said population that is not bound to a capture agent;
(d) selectively releasing one dissociably linked target subset, among said bound subsets, from said collection surface by dissociating said one dissociably linked subset from all or part of its solid support;
(e) isolating said selectively released target subset;
and
(f) dissociating the receptor of the other of said first or at least one other capture agent from its solid support;
and
(g) isolating the subset that had been bound to said other capture agent.

6 more elements

Dependencies are inferred from the claim text.

Facts

Application: 08/395,967Filed 28 Feb 1995 · continuation in part
Priority: 25 Mar 1991
Granted: 8 Jul 1997
Term ends: Not estimatedThe source records don’t support an estimate.

Classification15 codes

A23B 2/60
Preservation of foods or foodstuffs, in general: by treatment with electric currents without heating effect
B03C 1/035
Acting directly on the substance being separated: open gradient magnetic separators, i.e. separators in which the gap is unobstructed,…
G01N 33/54326
The carrier being characterised by its particulate form: magnetic particles
G01N 33/569
Immunoassay; Biospecific binding assay; Materials therefor: for microorganisms, e.g. protozoa, bacteria, viruses

Family

Serial674,678Parent of this continuation-in-part · filed 25 Mar 1991
Serial06,071Parent of this continuation-in-part · filed 15 Jan 1993
US 5,466,574 AParent of this continuation-in-part
US 5,186,827 AParent of this continuation-in-part
US 5,646,001 AThis patent

From the US filing history. Foreign family members aren’t listed.