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WO1989000693A1 - Protection de reactifs proteinaces - Google Patents

Protection de reactifs proteinaces Download PDF

Info

Publication number
WO1989000693A1
WO1989000693A1 PCT/GB1988/000586 GB8800586W WO8900693A1 WO 1989000693 A1 WO1989000693 A1 WO 1989000693A1 GB 8800586 W GB8800586 W GB 8800586W WO 8900693 A1 WO8900693 A1 WO 8900693A1
Authority
WO
WIPO (PCT)
Prior art keywords
reagent
substrate
solution
bound
trehalose
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.)
Ceased
Application number
PCT/GB1988/000586
Other languages
English (en)
Inventor
Donald Tills
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.)
FRESENIUS Ltd
Original Assignee
FRESENIUS Ltd
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 FRESENIUS Ltd filed Critical FRESENIUS Ltd
Publication of WO1989000693A1 publication Critical patent/WO1989000693A1/fr
Anticipated expiration legal-status Critical
Ceased 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
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding

Definitions

  • This invention concerns the protection or stabilisation of a proteinaceous reagent, for example an antibody, an antigen, or an enzyme, bound to a solid support, such as a plastics material. It is particularly applicable to anti ⁇ bodies bound to substrates such as the wells of microtitre plates for use in solid-phase assays for a wide range of antigens such as red or white blood cell antigens and bacterial and viral antigens .
  • Dried proteinaceous reagents tend to deteriorate with loss of activity and it is an object of this invention to stabilise such reagants so that they retain their activity on prolonged storage.
  • dried proteinaceous reagents can be effectively stabilised by means of the sugar trehalose.
  • the solution of trehalose is preferably aqueous and is preferably at a concentration of 2% .
  • the reagent forms a chemical bond with the plastics substrate, which is preferably polystyrene or most preferably polyvinylchloride , for example in the form of a multi-well microtitre plate.
  • the plastics substrate which is preferably polystyrene or most preferably polyvinylchloride , for example in the form of a multi-well microtitre plate.
  • a suitable coating buffer usually phosphate-based
  • pH usually phosphate-based
  • the reagen ⁇ is applied to the substrate in liquid form and may be bound to it by incubation, preferably at a temperature from about 18°C to 3 ⁇ °C. Any remaining liquid reagent is then discarded, and the substrate washed thoroughly with a washing solution that will both remove any unbound reagent and also act as a blocking buffer to prevent the substrate from binding any other reagent (e.g. antibody) that it might encounter.
  • the washing solution is then removed and the plate dried. Finally, the dried substrate is coated with a 2% solution of trehalose in distilled water and this is allowed to dry at a temperature between about 18°C and 37°C.
  • Substrates such as microtitration plates, prepared in this way can be stored for at least six months at 37°C in an aluminium foil pack containing a dessicant with the reagent remaining stable and active.
  • a U-bottomed microtitre plate of plastics material is coated with anti-A, anti-B, anti-A,B and anti- Rh(D) antibodies by applying a solution of the appropriate antibody to the wells of the plate, incubating the plate at 37°C, washing the plate several times with a "Tween” (Reg. Trade Mark for a surfactant which is a polyoxyethylene derivative of a fatty acid partial ester of a sorbitol anhydride) /
  • albumin/phosphate buffered saline washing solution dried by shaking and then by banging it on to absorbent paper.
  • a 2% solution of trehalose (50 ul per well) is then applied to the plate and the plate dried at 37°C.
  • the antibodies used can either be monoclonal or polyclonal antibodies which have been affinity-puri ied.
  • the thus-prepared plate may be stored for at least six months in an aluminium foil pack containing a dessleant.
  • the coated microtitre plate is removed from its aluminium foil container.
  • a 0.25% suspension of the erthrocytes .under test is made up and 50 ul added to each of the appropriate wells of the plate. If required the erythrocytes can be pre-treated with an enzyme.
  • microtitre plates are then centrifuged and the results read over an illuminated mirror.
  • the cells bind to the antibodies fixed t > the plates and so become evenly distributed over the bottom of the plate.
  • negatives all the cells form a single button at the bottom of the plate. Reading can be made easier by holding the plates at an angle of 80° for two minutes when only the negatives, where the blood cells remain unbound to the plate, will stream.

Landscapes

  • Health & Medical Sciences (AREA)
  • Immunology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Chemical & Material Sciences (AREA)
  • Urology & Nephrology (AREA)
  • Hematology (AREA)
  • Biomedical Technology (AREA)
  • Molecular Biology (AREA)
  • Medicinal Chemistry (AREA)
  • Analytical Chemistry (AREA)
  • Cell Biology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Food Science & Technology (AREA)
  • Biotechnology (AREA)
  • Physics & Mathematics (AREA)
  • Microbiology (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • General Physics & Mathematics (AREA)
  • Pathology (AREA)
  • Sampling And Sample Adjustment (AREA)
  • Agricultural Chemicals And Associated Chemicals (AREA)
  • Immobilizing And Processing Of Enzymes And Microorganisms (AREA)

Abstract

Cette invention se rapporte à la protection ou stabilisation d'un réactif protéinacé, par exemple un anticorps, un antigène, ou une enzyme, lié à un support solide tel qu'une matière plastique, et décrit un procédé permettant de stabiliser un réactif protéinacé séché lié à un substrat en matière plastique, par application sur celui-ci d'une solution de tréhalose que l'on laisse sécher sur ledit substrat.
PCT/GB1988/000586 1987-07-16 1988-07-18 Protection de reactifs proteinaces Ceased WO1989000693A1 (fr)

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
GB8716826 1987-07-16
GB878716826A GB8716826D0 (en) 1987-07-16 1987-07-16 Protection of proteinaceous reagents

Publications (1)

Publication Number Publication Date
WO1989000693A1 true WO1989000693A1 (fr) 1989-01-26

Family

ID=10620783

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/GB1988/000586 Ceased WO1989000693A1 (fr) 1987-07-16 1988-07-18 Protection de reactifs proteinaces

Country Status (3)

Country Link
AU (1) AU2080488A (fr)
GB (1) GB8716826D0 (fr)
WO (1) WO1989000693A1 (fr)

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0368624A3 (fr) * 1988-11-09 1991-06-05 Biotrack, Inc. Procédé et dispositif pour stabiliser et solubiliser des réactifs au latex
EP0897989A3 (fr) * 1997-08-15 2002-06-26 Becton, Dickinson and Company Excipient pour délivrer les particules à un échantillon
DE10252239A1 (de) * 2002-11-07 2004-05-27 Micronas Holding Gmbh Verfahren zum Drucken von Biomolekülen
WO2008055080A3 (fr) * 2006-10-31 2008-06-19 Sru Biosystems Inc Procédé pour le blocage d'une liaison à une protéine non spécifique sur une surface fonctionnalisée

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2016687A (en) * 1978-03-20 1979-09-26 Abbott Lab Sugar Coated Reagents for Solid Phase Immunoassay
EP0140489A1 (fr) * 1983-08-05 1985-05-08 Wako Pure Chemical Industries, Ltd. Méthode pour la stabilisation d'une substance immunologiquement active immobilisée sur un support insoluble et son utilisation dans la préparation d'un réactif pour la mesure de substances physiologiquement actives
JPS60149972A (ja) * 1984-01-17 1985-08-07 Yatoron:Kk 酵素標識抗体の安定化法
EP0192320A1 (fr) * 1985-01-11 1986-08-27 Unilever Plc Préparation de réactifs
WO1987000196A1 (fr) * 1985-07-09 1987-01-15 Quadrant Bioresources Limited Protection des proteines et similaires

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
GB2016687A (en) * 1978-03-20 1979-09-26 Abbott Lab Sugar Coated Reagents for Solid Phase Immunoassay
EP0140489A1 (fr) * 1983-08-05 1985-05-08 Wako Pure Chemical Industries, Ltd. Méthode pour la stabilisation d'une substance immunologiquement active immobilisée sur un support insoluble et son utilisation dans la préparation d'un réactif pour la mesure de substances physiologiquement actives
JPS60149972A (ja) * 1984-01-17 1985-08-07 Yatoron:Kk 酵素標識抗体の安定化法
EP0192320A1 (fr) * 1985-01-11 1986-08-27 Unilever Plc Préparation de réactifs
WO1987000196A1 (fr) * 1985-07-09 1987-01-15 Quadrant Bioresources Limited Protection des proteines et similaires

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
CHEMICAL ABSTRACTS, Vol. 103, 1985, Abstract 213222f; & JP,A,60 149 972 (7 Aug. 1985). *

Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0368624A3 (fr) * 1988-11-09 1991-06-05 Biotrack, Inc. Procédé et dispositif pour stabiliser et solubiliser des réactifs au latex
EP0897989A3 (fr) * 1997-08-15 2002-06-26 Becton, Dickinson and Company Excipient pour délivrer les particules à un échantillon
DE10252239A1 (de) * 2002-11-07 2004-05-27 Micronas Holding Gmbh Verfahren zum Drucken von Biomolekülen
DE10252239B4 (de) * 2002-11-07 2006-08-17 Micronas Holding Gmbh Verfahren zum Drucken von Biomolekülen
WO2008055080A3 (fr) * 2006-10-31 2008-06-19 Sru Biosystems Inc Procédé pour le blocage d'une liaison à une protéine non spécifique sur une surface fonctionnalisée

Also Published As

Publication number Publication date
GB8716826D0 (en) 1987-08-19
AU2080488A (en) 1989-02-13

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