Biologically-active polymers

Details Biologically-active polymers Biologically-active polymers

1998-02-13

2001-02-06

MacMillan, Keith D. (Department: 1618)

Chemistry: molecular biology and microbiology

Apparatus

Including measuring or testing

C435S007200, C436S517000, C436S518000, C436S164000, C436S172000, C424S048000, C204S418000

Reexamination Certificate

active

06184030

ABSTRACT:

TECHNICAL FIELD
This invention relates to biologically-active polymers that are useful for analyte detection and isolation and delivery of substances. More specifically, this invention relates to polymers that are capable of specifically and reversibly binding to analytes, including molecules and cells. The polymers of this invention are also capable of releasing substances upon electrical stimulation. The present invention relates to polymer membranes that act as substrates for cell adhesion, growth and differentiation. The present invention also relates to methods for making these polymer membranes and for specifically making polymer membranes and controlling them so as to have characteristic and specific properties for cell adhesion, growth and differentiation. These polymers have many applications in biological and chemical fields.
BACKGROUND OF THE INVENTION
Analyte Detection
For many years, scientists and physicians have sought rapid, simple and reliable methods for isolating and measuring analytes of all kinds including cells, cell organelles, molecules, and atoms. Most methods of isolating and measuring analytes have involved complicated and expensive analytical equipment such as spectrophotometers, densitometers, gamma and scintillation counters, flow cytometric devices, gas and ion exchange chromatographs, affinity columns, high performance liquid chromatographs and the like. In addition, many of these methods involve the use of various kinds of radiolabels such as
125
I,
3
H,
14
C,
32
P and others. These isotopes are expensive, require careful disposal and in some cases pose a health threat to laboratory personnel. Non-radioactive methods of analyte detection involve fluorescent, calorimetric, magnetic, and enzyme-linked assay methods among others. All of these methods involve expensive and sophisticated equipment, multiple additions of various reagents, mixing, incubating at different temperatures, and some separation steps, for example centrifugation, and take several hours if not days to complete. Furthermore, these methods do not permit in vivo measurement of an analyte or measurement of multiple analytes in a single system. Usually, a sample must be removed from a patient and subjected to various processing steps before attempting to include the sample in the assay system.
Accordingly what is needed is a rapid, simple, non-isotopic method that does not involve labeling of analytes or other reagents, and minimizes sample manipulation before performing the assay. Furthermore methods are needed that provide a means to measure one or more than one analyte in vivo and in vitro. What is also needed is a system that learns to detect patterns of analyte values associated with the presence of a certain condition or disease.
Substance Delivery
Health care providers such as physicians, nurses, therapists and others, and research scientists have long sought simple reliable, and controlled methods of delivering drugs and other substances to specific locations. Such targeted substance delivery has often been hampered by the typical problems involving substance solubility and stability, the rate and pattern of release of the substance, dilution in the general circulation before reaching the target site, and toxicity of the substance in systems that are not the desired target site.
What is desirable is a system that provides for targeted substance delivery in a controlled fashion. Such a system should be capable of delivering one or more than one substance, of being implanted into a patient, and of being activated by external signals to release substances at specific times and in specific amounts.
Analyte Detection and Substance Delivery System
What is also needed is a system that detects an analyte, analyzes the amount of analyte, and if the amount exceeds or does not meet a certain threshold, sends a signal to another membrane containing a desired substance for release of the substance at a desired location such as into a patient.
Isolation of Cells and Organelles
The human hematopoietic system is populated by cells of several different lineages. These “blood cells” may appear in bone marrow, the thymus, lymphatic tissue(s) and in blood such as umbilical cord blood, and also arterial blood and venous blood obtained centrally or peripherally. Within any specific lineage, there are a number of maturational stages. In most instances, the more immature developmental stages occur within bone marrow while the more mature and final stages of development occur in peripheral blood.
There are two major lineages: The myeloid lineage which matures into red blood cells, granulocytes, monocytes and megakaryocytes; and the lymphoid lineage which matures into B lymphocytes and T lymphocytes. Within each lineage and between each lineage, antigens are expressed differentially on the surface and in the cytoplasm of the cells in a given lineage. The expression of one or more antigens and/or the intensity of expression can be used to distinguish between maturational stages within a lineage and between lineages. Assignment of cell to lineage and to a maturational stage within a cell lineage indicates lineage commitment.
There are cells, however, which are uncommitted to any lineage (i.e., “progenitor” cells) and which, therefore, retain the ability to differentiate into each lineage. These undifferentiated, pluripotent progenitor cells will hereinafter be referred to as the “stem cells.” Therefore, all of mammalian hematopoietic cells can, in theory, be derived from a single stem cell. The stem cell is able to self-renew, so as to maintain a continuous source of pluripotent cells. In addition, when subject to particular environments and/or factors, the stem cells may differentiate to yield dedicated progenitor cells, which in turn may serve as the ancestor cells to a limited number of blood cell types. These ancestor cells will go through a number of stages before ultimately yielding a mature cell. Stem cells may be related to the lympho-hematopoietic system. Other stem cells may be unrelated to the lympho-hematopoietic system and may be derived from ectoderm, mesoderm, and endoderm. These stem cells may include germ cells such as, but not limited to, oogonia and spermatogonia, as well as myoblasts, fibroblasts, osteoblasts and neuroblasts.
The benefit of obtaining a pure population of stem cells is most readily recognized in the field of gene therapy. Briefly, gene therapy can be used to treat specific diseases caused by a defect in a particular gene. For example, sickle cell anemia is caused by a defect in a single gene. The cellular precursors of erythrocytes of sickle cell patients contain this defective gene which, in turn, codes for a defective form of the protein hemoglobin. The defective form results in the clinical condition of sickle cell anemia. Sickle cell anemia cannot be “cured” by conventional drug therapies because the underlying defect is in the gene which is included within every cell. Gene therapy seeks to replace or repopulate the cells of the hematopoietic system with cells that do not contain the defective gene but instead contain a “normal” gene. Using conventional recombinant DNA techniques, a “normal” gene is isolated, placed into a viral vector, and the viral vector is transfected into a cell capable of expressing the product coded for by the gene. The cell then must be introduced into the patient. If the “normal” gene product is produced, the patient is “cured” of the condition.
Bone marrow transplantation is an effective therapy for an increasing number of diseases. Graft Versus Host Disease (GVHD), however, limits bone marrow transplantation to recipients with histocompatibility (HLA)-matched sibling donors. Even then, approximately half of the allogenic bone marrow transplantation recipients develop GVHD. Current therapy for GVHD is imperfect and the disease can be disfiguring and/or lethal. Thus, risk of GVHD restricts the use of bone marrow transplantation to patients with otherwise fatal diseases, such as malignancies, severe aplastic anemia, and congenital i

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