Method of making a membrane having hydrophilic and hydrophobic surfaces for adhering cells or antibodies by using atomic oxygen or hydroxyl radicals

Claim 2

An organic polymer membrane having first and second opposing surfaces, wherein said first surface is hydrophilic and has an adjacent absorbent layer and said second surface is hydrophobic and contains adhered antibodies, said hydrophilic surface is prepared by a method comprising: exposing a hydrophobic exterior surface of said membrane to a dosage of atomic oxygen or hydroxyl radicals at a temperature below about 40° C., said dosage being sufficient to form said hydrophilic surface having a uniform surface layer of hydrophilic functional hydroxyl groups, wherein said dosage is generated by a flowing afterglow microwave discharge and said surface is outside of a plasma produced by said discharge, wherein said surface exposed to said dosage has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage.

Claim 3

An organic polymer membrane having a surface comprised of a plurality of hydrophilic regions bounded by adjacent hydrophobic regions, said hydrophilic regions containing adhered cells at a maximum density of one cell per each hydrophilic region and said hydrophobic regions are substantially free of adhered cells, said hydrophilic regions having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage.

Claim 4

An organic polymer membrane having a surface comprised of a plurality of hydrophilic regions bounded by adjacent hydrophobic regions, said hydrophilic regions containing adhered cells at a maximum density of one cell per each hydrophilic region and said hydrophobic regions are substantially free of adhered cells, said hydrophilic regions having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage.

Claim 5

The membrane of claim 2 wherein said hydrophilic regions are grafted with a compatibilizing compound selected from the group consisting of phosphorylcholines, peptides, lipids, proteins, nucleic acids, poly(ethylene oxide), poly(vinyl alcohol) and acyl terminated upper alkyl radicals.

Claim 6

The membrane of claim 3 wherein said compound is selected from the group consisting of one or more phosphorylcholines, peptides, lipids, proteins, and nucleic acids.

Claim 7

The membrane of claim 3, wherein said compatibilizing compound is selected from the group consisting of poly(vinyl alcohol) and poly(ethylene oxide).

Claim 8

The membrane of claim 2 in the form of a tube.

Claim 9

An assay method using a permeable organic polymer membrane having a hydrophilic surface formed opposite a hydrophobic surface, said hydrophilic surface having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, said assay comprising the steps of: (a) adhering known antibodies to said hydrophobic surface; (b) placing said hydrophilic surface in contact with an adjacent absorbent material; (c) covering said antibodies with a test solution, wherein antigens specific to said antibodies adhere thereto to form an antibody-antigen complex and non-adhered antigens migrate to said hydrophilic surface and are absorbed by said absorbent material; (d) covering said hydrophobic surface with a solution of said antibodies conjugated to an indicating enzyme, wherein said conjugated antibodies and said antigen-antibody complex form an antibody-antigen-antibody/enzyme complex and excess fluid is absorbed by said absorbent material; and (e) developing said antibody-antigen-antibody/enzyme complex with a solution of developing compound reactive with said enzyme to indicate presence of said antibody-antigen-antibody/enzyme complex as a positive result or an absence of said complexes as a negative result, wherein excess developing solution is absorbed by said absorbent material.

Claim 10

An assay method using a permeable organic polymer membrane having a hydrophilic surface formed opposite a hydrophobic surface, said hydrophilic surface having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, said assay comprising the steps of: (a) adhering known antibodies to said hydrophobic surface; (b) placing said hydrophilic surface in contact with an adjacent absorbent material; (c) covering said antibodies with a test solution, wherein antigens specific to said antibodies adhere thereto to form an antibody-antigen complex and non-adhered antigens migrate to said hydrophilic surface and are absorbed by said absorbent material; (d) covering said hydrophobic surface with a solution of said antibodies conjugated to an indicating enzyme, wherein said conjugated antibodies and said antigen-antibody complex form an antibody-antigen-antibody/enzyme complex and excess fluid is absorbed by said absorbent material; and (e) developing said antibody-antigen-antibody/enzyme complex with a solution of developing compound reactive with said enzyme to indicate presence of said antibody-antigen-antibody/enzyme complex as a positive result or an absence of said complexes as a negative result, wherein excess developing solution is absorbed by said absorbent material.

Claim 11

A cell culturing method using a permeable organic polymer membrane having a hydrophilic surface comprising a plurality of hydrophilic regions bounded by adjacent hydrophobic regions, said hydrophilic regions having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, said cell culturing method comprising the steps of: (a) distributing a group of cells on said membrane in a growth medium, wherein said cells adhere to said hydrophilic regions at a maximum density of one cell per each hydrophilic region and said hydrophobic regions are substantially free of adhered cells; (b) growing said cells on said membrane; and (c) obtaining a count of malignant cells growing on said membrane.

Claim 12

A cell culturing method using a permeable organic polymer membrane having a hydrophilic surface comprising a plurality of hydrophilic regions bounded by adjacent hydrophobic regions, said hydrophilic regions having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, said cell culturing method comprising the steps of: (a) distributing a group of cells on said membrane in a growth medium, wherein said cells adhere to said hydrophilic regions at a maximum density of one cell per each hydrophilic region and said hydrophobic regions are substantially free of adhered cells; (b) growing said cells on said membrane; and (c) obtaining a count of malignant cells growing on said membrane.

Claim 13

A cell culturing method using a permeable organic polymer membrane having first and second opposing surfaces, said first surface comprising a hydrophilic region bounded by a hydrophobic region, said hydrophilic region having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, second said cell culture method comprising the steps of: (a) forming cell culturing and product compartments on opposite sides of said membrane, wherein said first membrane surface is adjacent to said cell culturing compartment and said second membrane surface is adjacent to said product compartment, said cell culturing compartment comprising cells in a growth medium, said cells having a gene for expression of a desired product; (b) inducing said cells to adhere to said hydrophilic region of said first surface of said membrane, wherein said cells polarize with a basal cell wall adjacent said first surface; and (c) inducing migration of said nutrient from said nutrient compartment to said cells compartment by maintaining nutrient in said cell culturing compartment, wherein said cells absorb said nutrient and in response thereto secrete a product preferentially through said basal cell wall and said membrane into said product compartment.

Claim 14

A cell culturing method using a permeable organic polymer membrane having first and second opposing surfaces, said first surface comprising a hydrophilic region bounded by a hydrophobic region, said hydrophilic region having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, second said cell culture method comprising the steps of: (a) forming cell culturing and product compartments on opposite sides of said membrane, wherein said first membrane surface is adjacent to said cell culturing compartment and said second membrane surface is adjacent to said product compartment, said cell culturing compartment comprising cells in a growth medium, said cells having a gene for expression of a desired product; (b) inducing said cells to adhere to said hydrophilic region of said first surface of said membrane, wherein said cells polarize with a basal cell wall adjacent said first surface; and (c) inducing migration of said nutrient from said nutrient compartment to said cells compartment by maintaining nutrient in said cell culturing compartment, wherein said cells absorb said nutrient and in response thereto secrete a product preferentially through said basal cell wall and said membrane into said product compartment.

Claim 15

A cell culturing method using a permeable organic polymer membrane, said membrane having first and second opposing surfaces, said first surface comprising a hydrophilic surface and said second surface comprising a hydrophobic surface, said hydrophilic surface having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, said cell culture method comprising the steps of: (a) forming a nutrient compartment and a cells compartment on opposite sides of said membrane, wherein said first surface is adjacent said nutrient compartment and said second surface is adjacent to said cells compartment, said nutrient compartment comprising a solution of nutrient and said cells compartment comprising cells in a growth medium; and (b) inducing migration of said nutrient from said nutrient compartment to said cells compartment by maintaining a higher concentration of said nutrient in said nutrient compartment.

Claim 16

A cell culturing method using a permeable organic polymer membrane, said membrane having first and second opposing surfaces, said first surface comprising a hydrophilic surface and said second surface comprising a hydrophobic surface, said hydrophilic surface having a uniform surface layer of hydrophilic functional hydroxyl groups created by exposing an exterior hydrophobic surface of said membrane at a temperature below about 40° C. to a dosage of atomic oxygen or hydroxyl radicals, said dosage being generated by a flowing afterglow microwave discharge and said surface being outside of a plasma produced by said discharge, wherein said exterior surface exposed to said atomic oxygen has a depth of from about 50 to about 5000 angstroms and said dosage over said surface is within about 10 percent of an average dosage, said cell culture method comprising the steps of: (a) forming a nutrient compartment and a cells compartment on opposite sides of said membrane, wherein said first surface is adjacent said nutrient compartment and said second surface is adjacent to said cells compartment, said nutrient compartment comprising a solution of nutrient and said cells compartment comprising cells in a growth medium; and (b) inducing migration of said nutrient from said nutrient compartment to said cells compartment by maintaining a higher concentration of said nutrient in said nutrient compartment.