EP0233257A1 - Verfahren zur befestigung biologischer substanzen an festen substraten - Google Patents

Verfahren zur befestigung biologischer substanzen an festen substraten

Info

Publication number
EP0233257A1
EP0233257A1 EP86905147A EP86905147A EP0233257A1 EP 0233257 A1 EP0233257 A1 EP 0233257A1 EP 86905147 A EP86905147 A EP 86905147A EP 86905147 A EP86905147 A EP 86905147A EP 0233257 A1 EP0233257 A1 EP 0233257A1
Authority
EP
European Patent Office
Prior art keywords
biological substance
solid
substrate
group
attachment
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Withdrawn
Application number
EP86905147A
Other languages
English (en)
French (fr)
Other versions
EP0233257A4 (de
Inventor
Dinesh Kumar Sood
Ralph Edward Blandford Green
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
RMIT University
Melbourne Institute of Technology
Original Assignee
Royal Melbourne Institute of Technology Ltd
Melbourne Institute of Technology
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Royal Melbourne Institute of Technology Ltd, Melbourne Institute of Technology filed Critical Royal Melbourne Institute of Technology Ltd
Publication of EP0233257A1 publication Critical patent/EP0233257A1/de
Publication of EP0233257A4 publication Critical patent/EP0233257A4/de
Withdrawn legal-status Critical Current

Links

Classifications

    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
    • G01N33/545—Synthetic resin
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C59/00—Surface shaping of articles, e.g. embossing; Apparatus therefor
    • B29C59/16—Surface shaping of articles, e.g. embossing; Apparatus therefor by wave energy or particle radiation, e.g. infrared heating
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N11/00—Carrier-bound or immobilised enzymes; Carrier-bound or immobilised microbial cells; Preparation thereof
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/544—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being organic
    • G01N33/548—Carbohydrates, e.g. dextran
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/551—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals the carrier being inorganic
    • G01N33/552—Glass or silica
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29C—SHAPING OR JOINING OF PLASTICS; SHAPING OF MATERIAL IN A PLASTIC STATE, NOT OTHERWISE PROVIDED FOR; AFTER-TREATMENT OF THE SHAPED PRODUCTS, e.g. REPAIRING
    • B29C35/00—Heating, cooling or curing, e.g. crosslinking or vulcanising; Apparatus therefor
    • B29C35/02—Heating or curing, e.g. crosslinking or vulcanizing during moulding, e.g. in a mould
    • B29C35/08—Heating or curing, e.g. crosslinking or vulcanizing during moulding, e.g. in a mould by wave energy or particle radiation
    • B29C35/0866—Heating or curing, e.g. crosslinking or vulcanizing during moulding, e.g. in a mould by wave energy or particle radiation using particle radiation
    • B29C2035/0872—Heating or curing, e.g. crosslinking or vulcanizing during moulding, e.g. in a mould by wave energy or particle radiation using particle radiation using ion-radiation, e.g. alpha-rays
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B29—WORKING OF PLASTICS; WORKING OF SUBSTANCES IN A PLASTIC STATE IN GENERAL
    • B29L—INDEXING SCHEME ASSOCIATED WITH SUBCLASS B29C, RELATING TO PARTICULAR ARTICLES
    • B29L2031/00—Other particular articles
    • B29L2031/753—Medical equipment; Accessories therefor

Definitions

  • This invention relates to a method of attaching biological substances to solid substrates, and relates particularly, but not exclusively, to the attachment of biological materials such as antigens and/or antibodies to solid-phase substrates for use in . solid-phase immunoassays.
  • the invention also relates to the products which are produced by using the said method.
  • the attachment of one of the partners in an immunological reaction to a solid support facilitates the qualitative and/or quantitative measurement of the reaction.
  • the immobilization of one of the partners allows for easy separation of the reaction products from unreacted components.
  • the immobilised partner in the reaction captures its complementary reactant from solutions and binds it to a solid-phase. Once bound to this solid-phase its presence may then be measured either quantitatively or qualitatively.
  • Support materials used have included cellulose, agarose, dextran, polyacrylamide, glass, rubber and plastics including nylon, polystyrene and polyesters etc. Direct chemical bonding to produce covalent attachment may be used.
  • Chemical methods of attaching either an antigen or an antibody to the support material include the use of linking agents such as glutaraldehyde, cyanogen bromide, carbodiimides and aminoalkylsilanes. These techniques are well known to those skilled in the art.
  • Attachment of antigens or antibodies to plastic surfaces may be achieved by direct adsorption. This is a simple method of attachment however it has several disadvantages.
  • the adsorption of the antigen or antibody to the surface is achieved by hydrophobic interaction and van der aals forces. These are relatively weak attractive forces and as a result the attached material may elute from the surface during subsequent handling of the solid support (1) .
  • antigen attachment to such plates may be highly variable between different wells on the same plate (2) .
  • European patent No. 83111144 and European Patent No. 84103367 These methods involve preactivation with phenylalaninelysine copolymer and diphenylene-bis- diazonium compounds respectively.
  • a physical method of activating plastic microtitre plates is described in European Patent No. 82102257. This method entails the exposure of the microtitre plates to ⁇ -rays. Gamma irradiation produces better attachment (and also sterilization) due to ionization effects leading to the presence of some free radicals/bonds (3) .
  • soluble materials such as antibodies and antigens
  • Attachment may be facilitated using techniques known to those skilled in the art (4) . These techniques generally involve pre-treatment of the solid support with agents such as glutaraldehyde, poly-L-lysine, phenylalaninelysine copolymer or lectins derived from sources such as Glycine max, Limulus polyphemus, Lens culinaris and Phaseolus vulgaris, etc.
  • Cells may also be attached to a solid support by first attaching to the support an antibody which is specific for the cell. Attachment of the antibody may be achieved by the techniques previously mentioned.
  • a method of attaching a biological substance to a solid-phase substrate which comprises the steps of:- (i) subjecting a solid-phase substrate to ion bombardment, and (ii) contacting the treated substrate with a biological substance to be attached thereto.
  • the present invention also provides a product produced by the method described above.
  • the invention includes a solid-phase product comprising a solid-phase substrate, the surface of which has been treated by ion bombardment, and a biological substance attached to said treated surface.
  • Suitable biological materials for use in accordance with the present invention include protein materials such as antigens, antibodies, immunoglobulins such as IgG, IgM and IgE, and enzymes, as well as cellular material such as red blood cell membranes, whole cells such as bacteria and viruses, and hormones such as thyroid stimulating hormone (TSH) .
  • Suitable solid-phase substrates include in particular, but not exclusively, plastics materials such as polypropylene. Mylar, polystyrene, latex and Teflon; glass and silica; and cellulose and cellulose derivatives.
  • the step of ion bombardment of the solid-phase substrate may, for example, be carried out by bombarding the substrate with a beam of energetic Ar ions. Whilst the exact mechanism which leads to the strong specific attachment of biological substances to such a treated substrate is not well understood at present, it is believed that ion bombardment could produce one or more of the following effects which are conducive to strong bonding at the interface:
  • ion bombardment produces a much higher ionization density (stopping power) and can also produce atomic recoils, it is capable of generating metastable chemical surface states/compositions not attainable by milder forms of ionizing radiation or by any conventional equilibrium chemistry.
  • the ion bombardment technique is believed to be quite unique in that, it can not only produce effects similar to those generated by milder ionizing radiation, but also can produce many new metastable chemical states (dangling bonds) . This feature is believed to be responsible for more universal success of ion bombardment in producing enhanced attachment of many different types of biological materials to several different types of plastics.
  • the biological substance is then brought into contact with the treated substrate.
  • the unique bonding process of the present invention may be conducted at room temperature and does not require any extraneous chemical bonding agents. In this respect,
  • the method of the present invention is utilised to attach red blood cell membranes to transparent plastic solid-phase substrates chosen as test substrates, the first being polypropylene (which possesses no reactive groups) , and the second being Mylar (which has some reactive groups) .
  • red blood cell membranes in colloidal suspension in a suitable liquid are then injected in situ (in vacuum) onto the surface which has been treated by ion bombardment.
  • the liquid quickly spreads to a fine coating with the red blood cell membranes being bonded so strongly to the substrate surface that even extended ultrasonic treatment in water has no effect in removal of the membranes from the surface.
  • Red blood cell membranes were prepared from fresh anticoagulated blood by the process of calcium phosphate cosedimentation (8) .
  • Sodium azide was incorporated at a final concentration of 0.1% to inhibit bacterial contamination during the processing of the sample for attachment to the plastic substrate surface.
  • Polypropylene strips were treated with Ar ions in apparatus illustrated in Figure 1.
  • the strips were placed in sample holder 11 within the target chamber 10 with the system maintained totally under vacuum, and bboommbbaarrddeedd with accelerated Ar ions 12 through aperture plate 13.
  • the sample holder 11 was transferred through air lock valve 20 to an adjacent air lock 21 without breaking vacuum ( Figure 1).
  • air lock 21 a controlled volume of the red blood cell membrane calcium phosphate suspension was then injected onto the plastic strips through biological material injection port 22.
  • the liquid did not wet the unbombarded regions of the polypropylene and quickly spread to produce a coating on the bombarded regions.
  • the plastic strips were removed from the air lock and excess liquid removed. After allowing to air dry the strength of adhesion of the red cell membrane/calcium phosphate mixture was assessed by immersing the strip in water and subjecting it to ultrasonic cleaning for five minutes. The strips were dried in air and then examined by optical microscopy.
  • Figures 2a and 2b show the results for bombarded and unbombarded regions on polypropylene and Mylar respectively.
  • IgG was purified from human serum by
  • Sepharose-Staph A affinity column chromatography. An aliquot of the purified IgG was radiolabelled with 125I by the Chloramine T method. Radiolabelled IgG was added to the purified IgG as a tracer to give a ratio of labelled to unlabelled IgG of between 1:20 and 1:30.
  • This radiolabelled material was then used to examine the influence of IgG concentration, buffer pH and reaction time on the interaction of IgG with ion bombarded plastic surfaces.
  • IgG was diluted in phosphate buffer pH 7.0 and carbonate buffer pH 9.6 over a concentration range of lO ⁇ g/ml to lOO ⁇ g/ml. 40 ⁇ l samples were applied to wells (0.6cm diameter) impressed on ion bombarded and virgin plastic strips. Following incubation at room temperature the strips were rinsed with deionised water, the wells cut from the strip and the radioactivity due to adherent IgG counted in a ⁇ counter.
  • Table 1 indicates the influence of different forms of ion bombardment and different plastics on the uptake of IgG. This clearly shows that ion bombardment can increase the uptake of IgG to plastic surfaces and that the efficiency varies depending on the species of the ion employed and the type of plastic used.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Immunology (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Molecular Biology (AREA)
  • Biomedical Technology (AREA)
  • Hematology (AREA)
  • Urology & Nephrology (AREA)
  • General Health & Medical Sciences (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • Physics & Mathematics (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Food Science & Technology (AREA)
  • Cell Biology (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Genetics & Genomics (AREA)
  • Wood Science & Technology (AREA)
  • Zoology (AREA)
  • Organic Chemistry (AREA)
  • Toxicology (AREA)
  • Inorganic Chemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • Treatments Of Macromolecular Shaped Articles (AREA)
  • Medicines Containing Antibodies Or Antigens For Use As Internal Diagnostic Agents (AREA)
EP19860905147 1985-08-19 1986-08-18 Verfahren zur befestigung biologischer substanzen an festen substraten. Withdrawn EP0233257A4 (de)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
AUPH202785 1985-08-19
AU2027/85 1985-08-19

Publications (2)

Publication Number Publication Date
EP0233257A1 true EP0233257A1 (de) 1987-08-26
EP0233257A4 EP0233257A4 (de) 1989-01-26

Family

ID=3771228

Family Applications (1)

Application Number Title Priority Date Filing Date
EP19860905147 Withdrawn EP0233257A4 (de) 1985-08-19 1986-08-18 Verfahren zur befestigung biologischer substanzen an festen substraten.

Country Status (7)

Country Link
EP (1) EP0233257A4 (de)
JP (1) JPS63500893A (de)
BR (1) BR8606830A (de)
DK (1) DK200387D0 (de)
FI (1) FI871599A (de)
HU (1) HUT43092A (de)
WO (1) WO1987001120A1 (de)

Families Citing this family (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5055316A (en) * 1988-04-20 1991-10-08 Washington Research Foundation Tight binding of proteins to surfaces
US5389195A (en) * 1991-03-07 1995-02-14 Minnesota Mining And Manufacturing Company Surface modification by accelerated plasma or ions
DE19538523A1 (de) * 1995-10-06 1997-04-10 Helmut Prof Dr Kaeufer Biokompatible Kunststoffe, Verfahren zu deren Herstellung und Anwendungsgebiete
JP5952522B2 (ja) * 2008-03-31 2016-07-13 旭化成株式会社 セルロース誘導体微粒子、その分散液、その分散体及び診断薬

Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0104608A1 (de) * 1982-09-24 1984-04-04 Becton Dickinson and Company Chemisch modifizierte Oberfläche zur Befestigung grosser Moleküle

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE3111474A1 (de) * 1981-03-24 1982-10-07 Behringwerke Ag, 3550 Marburg "mittel zur immunologischen diagnose sowie verfahren zu seiner herstellung"

Patent Citations (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0104608A1 (de) * 1982-09-24 1984-04-04 Becton Dickinson and Company Chemisch modifizierte Oberfläche zur Befestigung grosser Moleküle

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
See also references of WO8701120A1 *

Also Published As

Publication number Publication date
BR8606830A (pt) 1987-10-27
JPS63500893A (ja) 1988-03-31
FI871599A0 (fi) 1987-04-13
HUT43092A (en) 1987-09-28
DK200387A (da) 1987-04-15
EP0233257A4 (de) 1989-01-26
FI871599A (fi) 1987-04-13
WO1987001120A1 (en) 1987-02-26
DK200387D0 (da) 1987-04-15

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Legal Events

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PUAI Public reference made under article 153(3) epc to a published international application that has entered the european phase

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Effective date: 19901115

RIN1 Information on inventor provided before grant (corrected)

Inventor name: GREEN, RALPH, EDWARD, BLANDFORD

Inventor name: SOOD, DINESH, KUMAR